isp-1

UniProt ID: O44512
Organism: Caenorhabditis elegans
Review Status: DRAFT
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Gene Description

isp-1 encodes the Rieske iron-sulfur protein (ISP), the [2Fe-2S]-cluster-bearing catalytic subunit of mitochondrial respiratory complex III (the cytochrome bc1 / ubiquinol-cytochrome c oxidoreductase complex, EC 7.1.1.8). Anchored in the mitochondrial inner membrane, its mobile Rieske head domain accepts an electron from ubiquinol at the complex III Qo site and delivers it to cytochrome c1, performing the electron-transfer step of the protonmotive Q-cycle and thereby feeding electrons into the respiratory chain. In C. elegans the partial loss-of-function allele isp-1(qm150) is a classic mitochondrial ("Mit") longevity mutant: it lowers oxygen consumption and extends lifespan, with the lifespan extension driven by an elevated mitochondrial superoxide signal rather than by reduced oxidative damage.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0016491 oxidoreductase activity
IBA
GO_REF:0000033
MODIFY
Summary: Correct but uninformatively general. ISP-1's specific molecular function is electron transfer through its Rieske [2Fe-2S] cluster; the complex-level oxidoreductase reaction is captured more precisely by GO:0008121.
Reason: oxidoreductase activity is a high-level parent. The subunit-specific activity of the Rieske protein is electron transfer (GO:0009055), enabled via its 2Fe-2S cluster; complex III's overall reaction is quinol-cytochrome-c reductase activity (GO:0008121), already separately annotated.
Propagation Review
Root cause: TERM SCOPING PROBLEM
Failure modes: GRANULARITY MISMATCH
Sources checked:
PANTHER:PTN000015358 · Rieske / complex III oxidoreductase node
IBA propagates the broad parent oxidoreductase activity; the correct subunit-level term is electron transfer activity (GO:0009055).
Proposed replacements: electron transfer activity
Supporting Evidence:
PMID:26504246
encodes the Rieske iron-sulfur protein subunit of cytochrome c oxidoreductase (complex III of the electron transport chain)
GO:0045275 respiratory chain complex III
IBA
GO_REF:0000033
ACCEPT
Summary: Core annotation. ISP-1 is the Rieske iron-sulfur subunit of respiratory chain complex III (cytochrome bc1).
Reason: Well established across eukaryotes and in C. elegans; ISP-1 is an integral catalytic subunit of complex III.
Supporting Evidence:
PMID:21151885
subunits of complex I and III of the mitochondrial respiratory chain
PMID:26504246
complex III of the electron transport chain
GO:0006122 mitochondrial electron transport, ubiquinol to cytochrome c
IBA
GO_REF:0000033
ACCEPT
Summary: Core biological process. Within complex III, ISP-1 transfers electrons from ubiquinol to cytochrome c1 (and onward to cytochrome c).
Reason: This is the pathway step complex III performs and to which the Rieske subunit is central; phylogenetically well supported and consistent with the experimental worm data.
Supporting Evidence:
PMID:21151885
Partial loss-of-function mutations in these genes decrease electron transport
file:worm/isp-1/isp-1-deep-research-falcon.md
transfers electrons from ubiquinol to cytochrome c1
GO:0005743 mitochondrial inner membrane
IEA
GO_REF:0000044
ACCEPT
Summary: Core localization. Complex III, including the Rieske subunit, resides in the mitochondrial inner membrane.
Reason: UniProt subcellular location; ISP-1 has a single-pass inner-membrane anchor with its Rieske head projecting to the intermembrane-space side.
Supporting Evidence:
file:worm/isp-1/isp-1-uniprot.txt
C:mitochondrial inner membrane; IEA:UniProtKB-SubCell
GO:0008121 quinol-cytochrome-c reductase activity
IEA
GO_REF:0000120
ACCEPT
Summary: Complex III catalytic activity (EC 7.1.1.8) to which ISP-1 contributes the essential electron-transfer step. Retained as a core molecular function of the subunit in the context of the assembled complex.
Reason: ISP-1 is a catalytic subunit of ubiquinol-cytochrome c oxidoreductase; the Rieske cluster carries out the electron-transfer half of this reaction. Best represented in core_functions as contributes_to (a complex-level activity).
Supporting Evidence:
file:worm/isp-1/isp-1-uniprot.txt
EC=7.1.1.8
PMID:26504246
subunit of cytochrome c oxidoreductase
GO:0016020 membrane
IEA
GO_REF:0000120
MARK AS OVER ANNOTATED
Summary: Uninformative general parent, redundant with the specific and correct mitochondrial inner membrane annotation (GO:0005743).
Reason: "membrane" adds no information beyond the more precise GO:0005743 already assigned; it is an over-general IEA byproduct.
GO:0051537 2 iron, 2 sulfur cluster binding
IEA
GO_REF:0000002
ACCEPT
Summary: Core molecular function. The defining feature of the Rieske protein: it coordinates a high-potential [2Fe-2S] cluster, the redox center used for electron transfer.
Reason: UniProt cofactor annotation and Rieske-family conservation; one [2Fe-2S] cluster is bound per subunit.
Supporting Evidence:
file:worm/isp-1/isp-1-uniprot.txt
Binds 1 [2Fe-2S] cluster per subunit
file:worm/isp-1/isp-1-uniprot.txt
The Rieske protein is a high potential 2Fe-2S protein
GO:1902600 proton transmembrane transport
IEA
GO_REF:0000108
KEEP AS NON CORE
Summary: Complex III couples electron transfer to proton translocation across the inner membrane via the Q-cycle. This is a complex-level chemiosmotic outcome, not ISP-1's direct molecular function (electron transfer); retained as non-core.
Reason: Proton translocation is an emergent property of the intact Q-cycle to which ISP-1 contributes, rather than an activity ISP-1 performs on its own. Correct to retain, but the subunit's core function is electron transfer.
GO:0006122 mitochondrial electron transport, ubiquinol to cytochrome c
IMP
PMID:16920626
Mitochondrial complex I function modulates volatile anesthet...
ACCEPT
Summary: Experimentally supported in C. elegans. The isp-1 complex III mutant shows impaired mitochondrial respiration/oxidative phosphorylation, directly implicating isp-1 in mitochondrial electron transport.
Reason: Falk et al. measured respiration in the isp-1 (complex III) mutant and found diminished complex II-dependent oxidative phosphorylation, consistent with a defect in the ubiquinol-to-cytochrome c step; core process, experimentally anchored.
Supporting Evidence:

Core Functions

Electron-transfer subunit of mitochondrial complex III: within the cytochrome bc1 complex, ISP-1's mobile Rieske head accepts an electron from ubiquinol at the Qo site and delivers it to cytochrome c1, the electron-transfer step of the ubiquinol-cytochrome c reductase (EC 7.1.1.8) reaction in the protonmotive Q-cycle.

Supporting Evidence:
  • PMID:26504246
    encodes the Rieske iron-sulfur protein subunit of cytochrome c oxidoreductase (complex III of the electron transport chain)
  • PMID:21151885
    subunits of complex I and III of the mitochondrial respiratory chain

Coordinates the Rieske high-potential [2Fe-2S] cluster (one per subunit), the redox cofactor that carries out the electron-transfer step of complex III.

Supporting Evidence:
  • file:worm/isp-1/isp-1-uniprot.txt
    Binds 1 [2Fe-2S] cluster per subunit

References

Gene Ontology annotation through association of InterPro records with GO terms
Annotation inferences using phylogenetic trees
Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping, accompanied by conservative changes to GO terms applied by UniProt
Automatic assignment of GO terms using logical inference, based on on inter-ontology links
Combined Automated Annotation using Multiple IEA Methods
Mitochondrial complex I function modulates volatile anesthetic sensitivity in C. elegans.
Mitochondrial electron transport is a key determinant of life span in Caenorhabditis elegans.
A mitochondrial superoxide signal triggers increased longevity in Caenorhabditis elegans.
Two modes of mitochondrial dysfunction lead independently to lifespan extension in Caenorhabditis elegans.
Tether mutations that restore function and suppress pleiotropic phenotypes of the C. elegans isp-1(qm150) Rieske iron-sulfur protein.
file:worm/isp-1/isp-1-uniprot.txt
UniProtKB O44512 (isp-1, C. elegans) - Cytochrome b-c1 complex subunit Rieske, mitochondrial
file:worm/isp-1/isp-1-deep-research-falcon.md
Falcon (Edison) deep research report for C. elegans isp-1

Knowledge Gaps

What is not known — curated, literature-grounded statements of the open unknowns (the inverse of core functions).

Gap: The molecular mechanism by which the isp-1(qm150) Rieske substitution is converted into the pro-longevity retrograde signal is undetermined. It is established that qm150 elevates mitochondrial superoxide and that this elevation is necessary and sufficient for the lifespan extension, but the causal chain from the specific perturbation of ISP-1 head/tether dynamics, through altered Qo-site electron transfer and superoxide production, to the downstream transcriptional longevity program has not been resolved.

OPEN BIOLOGY RESIDUAL_SUBGAP

What is known: Firmly established: isp-1 encodes the Rieske [2Fe-2S] subunit of complex III; qm150 is a partial loss-of-function allele that lowers oxygen consumption and extends lifespan; the longevity requires elevated mitochondrial superoxide (abolished by antioxidants, phenocopied by paraquat); and intragenic suppressors that restore function map to a conserved six-residue tether region ("spring-loaded" model). What is unresolved is the signal-generating step itself.

Significance: isp-1(qm150) is a canonical mitohormesis / Mit longevity model. Defining the signal-generating step would connect a specific electron-transport-chain lesion to a conserved lifespan-extension program with relevance to mitochondrial disease and aging.

What would resolve it: Structure-guided electron-transfer and superoxide measurements on qm150 and its intragenic tether-region suppressors, combined with epistasis to the retrograde transcriptional effectors, to identify the step that generates the superoxide longevity signal.

Provenance (the field's own admissions):

Deep Research

Falcon

(isp-1-deep-research-falcon.md)
Comprehensive Research Report: *isp-1* (F42G8.12) — Rieske Iron-Sulfur Protein of Mitochondrial Complex III in *Caenorhabditis elegans* Falcon Edison Scientific Literature 44 citations 2 artifacts 2026-07-04T14:48:30.379729

The research report should be a detailed narrative explaining the function, biological processes, and localization of the gene product. Citations should be given for all claims.

You should prioritize authoritative reviews and primary scientific literature when conducting research. You can supplement
this with annotations you find in gene/protein databases, but these can be outdated or inaccurate.

We are specifically interested in the primary function of the gene - for enzymes, what reaction is catalyzed, and what is the substrate specificity? For transporters, what is the substrate? For structural proteins or adapters, what is the broader structural role? For signaling molecules, what is the role in the pathway.

We are interested in where in or outside the cell the gene product carries out its function.

We are also interested in the signaling or biochemical pathways in which the gene functions. We are less interested in broad pleiotropic effects, except where these elucidate the precise role.

Include evidence where possible. We are interested in both experimental evidence as well as inference from structure, evolution, or bioinformatic analysis. Precise studies should be prioritized over high-throughput, where available.

Comprehensive Research Report: isp-1 (F42G8.12) — Rieske Iron-Sulfur Protein of Mitochondrial Complex III in Caenorhabditis elegans

Gene: isp-1 | UniProt: O44512 | Organism: Caenorhabditis elegans | Protein: Cytochrome b-c1 complex subunit Rieske, mitochondrial | EC: 7.1.1.8


1. Gene Identity and Protein Overview

The isp-1 gene (locus F42G8.12) in C. elegans encodes the Rieske iron-sulfur protein (ISP), a catalytic subunit of mitochondrial Complex III (ubiquinol:cytochrome c oxidoreductase, also known as the cytochrome bc1 complex) (jafari2016newfunctionaland pages 1-4). ISP-1 belongs to the highly conserved Rieske iron-sulfur protein family, with orthologs including UQCRFS1 in mammals (osz2025mutationsofthe pages 4-6). The protein contains characteristic domains including a Rieske 2Fe-2S cluster domain (IPR017941), a Rieske 2Fe-2S superfamily domain (IPR036922), a Rieske Fe-S protein domain (IPR014349), and a bc1 Rieske transmembrane superfamily domain (IPR037008), all consistent with its function as a catalytic electron transfer component within the Q-cycle of Complex III.


2. Protein Structure and Domain Architecture

ISP-1 comprises three structurally and functionally distinct domains that are essential for its role in the Q-cycle mechanism of Complex III:

Domain/Feature Description Function
Transmembrane helix (anchor) Single N-terminal transmembrane α-helix that anchors ISP-1 in the mitochondrial inner membrane as a core subunit of Complex III/cytochrome bc1. The membrane anchor remains relatively static while the catalytic domain moves. (jafari2016newfunctionaland pages 1-4, yang2012rieskeiron–sulfurprotein pages 1-3) Positions ISP-1 within Complex III and maintains the spatial framework needed for electron transfer between quinol oxidation and cytochrome c1 reduction. (jafari2016newfunctionaland pages 1-4, yang2012rieskeiron–sulfurprotein pages 1-3)
Tether domain (spring mechanism) Flexible linker connecting the membrane anchor to the extrinsic catalytic head. In mechanistic models it behaves as a chemical “spring,” alternating between extended and relaxed/helical conformations during catalysis; the qm150 Pro→Ser mutation lies in this region. (jafari2016newfunctionaland pages 1-4, jafari2016newfunctionaland pages 10-16, jafari2016newfunctionaland pages 6-8, jafari2016newfunctionaland pages 4-6) Enables controlled movement of the head domain between the Qo site and cytochrome c1, thereby regulating enzyme-substrate complex formation, electron flux through the Q-cycle, and ROS propensity. (jafari2016newfunctionaland pages 10-16, jafari2016newfunctionaland pages 6-8, jafari2016newfunctionaland pages 4-6)
Head domain with 2Fe-2S cluster Extrinsic/mobile catalytic head domain containing the Rieske 2Fe-2S cluster with distinctive histidine coordination and relatively high redox potential. This domain projects to the P side/intermembrane-space side of the membrane. (jafari2016newfunctionaland pages 1-4, schmidt2004rieskeiron–sulfurproteins pages 1-2) Accepts an electron from ubiquinol at the Qo site and later donates that electron to heme c1; its mobility and redox chemistry are central to Complex III catalysis. (jafari2016newfunctionaland pages 1-4, jafari2016newfunctionaland pages 10-16, schmidt2004rieskeiron–sulfurproteins pages 1-2)
Qo site interaction ISP-1 docks at the quinol oxidation (Qo) site near cytochrome b, where oxidized ISP interacts with ubiquinol (QH2). The first electron-transfer step reduces the 2Fe-2S cluster and generates a semiquinone intermediate. (jafari2016newfunctionaland pages 10-16, jafari2016newfunctionaland pages 6-8) Executes the initial oxidation of QH2 and bifurcates electron flow in the Q-cycle, a key energy-conserving step in Complex III. (jafari2016newfunctionaland pages 10-16, jafari2016newfunctionaland pages 6-8, gurung2005theironsulfurcluster pages 1-1)
Cytochrome c1 interaction After reduction at the Qo site, the ISP-1 head swings toward cytochrome c1 into the c1-state conformation, where the reduced 2Fe-2S center transfers its electron to heme c1. (jafari2016newfunctionaland pages 1-4, jafari2016newfunctionaland pages 10-16, yang2012rieskeiron–sulfurprotein pages 1-3, crofts1999physicochemicalaspectsof pages 11-12) Couples quinol oxidation to reduction of cytochrome c1 and ultimately cytochrome c, supporting downstream electron flow to Complex IV. (jafari2016newfunctionaland pages 1-4, yang2012rieskeiron–sulfurprotein pages 1-3)
Proton-exiting gate role Experimental disruption of the Rieske 2Fe-2S center creates a proton leak, supporting the model that the cluster/head region acts as a proton-exiting gate in cytochrome bc1. (gurung2005theironsulfurcluster pages 1-1, gurung2005theironsulfurcluster pages 5-5) Prevents nonspecific proton leakage and helps couple electron transfer to proton translocation, preserving the proton motive force used for ATP synthesis. (gurung2005theironsulfurcluster pages 1-1, gurung2005theironsulfurcluster pages 5-5)
Overall Q-cycle reaction (EC 7.1.1.8; ubiquinol:ferricytochrome-c reductase) ISP-1 is a catalytic subunit of Complex III/ubiquinol:cytochrome c oxidoreductase. In the Q-cycle, it transfers electrons from ubiquinol to cytochrome c1 while coordinating with the Qo and Qi sites to couple redox chemistry to proton translocation across the inner membrane. (jafari2016newfunctionaland pages 1-4, jafari2016newfunctionaland pages 10-16, gurung2005theironsulfurcluster pages 1-1, yang2012rieskeiron–sulfurprotein pages 1-3) Contributes to the net reaction of ubiquinol oxidation and cytochrome c reduction while generating the proton gradient required for oxidative phosphorylation and ATP production. (jafari2016newfunctionaland pages 1-4, jafari2016newfunctionaland pages 10-16, yang2012rieskeiron–sulfurprotein pages 1-3)

Table: This table summarizes the major structural features of the C. elegans ISP-1/Rieske iron-sulfur protein and links each feature to its role in Complex III catalysis. It is useful for connecting protein architecture to the Q-cycle mechanism and the gene’s primary biochemical function.

Transmembrane anchor. A single N-terminal transmembrane α-helix anchors ISP-1 in the mitochondrial inner membrane, positioning it as a core subunit of the cytochrome bc1 complex (jafari2016newfunctionaland pages 1-4, yang2012rieskeiron–sulfurprotein pages 1-3). This anchor remains relatively static during catalysis.

Tether domain. A flexible linker region connects the membrane anchor to the catalytic head domain. This tether operates as a chemical "spring," alternating between extended and relaxed (helical) conformations to enable the large-scale domain movement required for catalysis (jafari2016newfunctionaland pages 1-4, jafari2016newfunctionaland pages 10-16). The tether contains conserved amino acids critical for mediating interactions between cytochrome b and the ISP head domain (jafari2016newfunctionaland pages 6-8). Notably, the well-characterized isp-1(qm150) mutation—a proline-to-serine substitution—maps to this tether region and profoundly affects protein function (jafari2016newfunctionaland pages 6-8, jafari2016newfunctionaland pages 4-6).

Extrinsic head domain with 2Fe-2S cluster. The catalytic head domain projects into the intermembrane space (P-side) and harbors the Rieske [2Fe-2S] cluster with distinctive histidine-coordinated nitrogen ligands, conferring a relatively high redox potential compared to other iron-sulfur centers (schmidt2004rieskeiron–sulfurproteins pages 1-2). This domain undergoes large-scale movement of approximately 16–22 Å between the Qo site on cytochrome b and cytochrome c1, with root-mean-square displacement occurring in less than 25 nanoseconds (crofts1999physicochemicalaspectsof pages 11-12).


3. Primary Enzymatic Function: The Q-Cycle Mechanism

ISP-1 functions as a catalytic subunit of Complex III, which catalyzes the oxidation of ubiquinol (coenzyme QH₂) coupled to the reduction of cytochrome c and the translocation of protons across the mitochondrial inner membrane (EC 7.1.1.8) (jafari2016newfunctionaland pages 1-4, gurung2005theironsulfurcluster pages 1-1). The specific role of ISP-1 within the Q-cycle is as follows:

Step 1 — Ubiquinol oxidation at the Qo site. The oxidized 2Fe-2S cluster (ISP_ox) of the head domain docks at the Qo site (quinol oxidation site) near cytochrome b, where it accepts one electron from bound ubiquinol (QH₂). This first electron transfer reduces ISP_ox to ISP_H and generates a transient semiquinone intermediate (SQo) (jafari2016newfunctionaland pages 10-16).

Step 2 — Electron shuttle to cytochrome c1. The reduced ISP head dissociates from the Qo site and swings on its tether to dock at cytochrome c1, where the electron is transferred from the reduced 2Fe-2S cluster to heme c1, coupled with release of a proton to the P-phase (intermembrane space). This constitutes the "high-potential chain" of electron flow, ultimately reducing soluble cytochrome c (jafari2016newfunctionaland pages 10-16, yang2012rieskeiron–sulfurprotein pages 1-3).

Step 3 — Bifurcation of electron flow. The second electron from ubiquinol oxidation follows the separate "low-potential chain" through heme b_L and heme b_H of cytochrome b to the Qi site (quinone reduction site), where ubiquinone is reduced to ubiquinol. This bifurcated electron transfer is fundamental to the Q-cycle's proton-translocation stoichiometry of 2H⁺/e⁻ (jafari2016newfunctionaland pages 10-16, osz2025mutationsofthe pages 19-21, gurung2005theironsulfurcluster pages 1-1).

Proton-exiting gate function. Experimental work has demonstrated that the 2Fe-2S cluster also serves as a proton-exiting gate: destruction or genetic elimination of the cluster creates a proton-leaking channel in the bc1 complex, abolishing proton-pumping activity. This indicates the ISP head domain is critical for maintaining the integrity of proton translocation coupled to electron transfer (gurung2005theironsulfurcluster pages 1-1, gurung2005theironsulfurcluster pages 5-5).


4. Subcellular Localization

ISP-1 is localized to the mitochondrial inner membrane, where it functions as an integral component of the energy-conserving electron transport chain (jafari2016newfunctionaland pages 1-4, yang2012rieskeiron–sulfurprotein pages 1-3, gurung2005theironsulfurcluster pages 1-1). The protein's topology places the transmembrane anchor within the lipid bilayer while the catalytic head domain with the 2Fe-2S cluster extends into the intermembrane space (P-side) (jafari2016newfunctionaland pages 1-4, yang2012rieskeiron–sulfurprotein pages 1-3). ISP-1 also plays a structural role in stabilizing higher-order respiratory supercomplexes (I:III:IV respirasomes), which are proposed to increase electron transport chain efficiency and reduce ROS production (osz2025mutationsofthe pages 19-21).


5. The isp-1(qm150) Mutation: Functional Insights from Genetic Analysis

The best-characterized allele of isp-1 is qm150, a proline-to-serine substitution in the tether domain that inhibits electron flux into the Qo site by interfering with formation of the enzyme-substrate complex between QH₂ and ISP_ox (jafari2016newfunctionaland pages 6-8, jafari2016newfunctionaland pages 4-6). This mutation results in drastically diminished respiratory function and causes pleiotropic phenotypes including:

  • Extended lifespanisp-1(qm150) mutants are significantly long-lived compared to wild-type animals (jafari2016newfunctionaland pages 1-4, lee2010inhibitionofrespiration pages 2-3)
  • Slow development — delayed embryonic and postembryonic development (osz2025mutationsofthe pages 19-21, jafari2016newfunctionaland pages 1-4)
  • Reduced brood size and fecundity (osz2025mutationsofthe pages 19-21, jafari2016newfunctionaland pages 1-4)
  • Small body size (jafari2016newfunctionaland pages 1-4)
  • Decreased movement and pharyngeal pumping rate (jafari2016newfunctionaland pages 1-4)
  • Sensitivity to hyperoxia — inability to develop past the L2 larval stage under 100% O₂ (jafari2016newfunctionaland pages 4-6)
  • Elevated reactive oxygen species (ROS) — measured by DCF-DA fluorescence and DHE sensors (lee2010inhibitionofrespiration pages 2-3, lee2010inhibitionofrespiration pages 4-5)
  • Disruption of I:III:IV supercomplexes and impaired complex I activity (osz2025mutationsofthe pages 19-21)

Intragenic suppressor mutations identified in the tether region partially restore electron transfer rates and suppress some pleiotropic phenotypes, but at the cost of increased ROS production, demonstrating the intimate relationship between tether mechanics and Q-cycle gating (jafari2016newfunctionaland pages 4-6, jafari2016newfunctionaland pages 6-8).


6. Signaling and Biochemical Pathways Downstream of ISP-1 Dysfunction

Mild mitochondrial dysfunction caused by isp-1 mutation activates multiple conserved stress response and longevity-promoting signaling pathways. The central upstream signal appears to be elevated mitochondrial ROS, which triggers several downstream transcriptional programs.

Pathway/Transcription Factor Role in isp-1 Longevity Key Evidence Key Reference
HIF-1 Required for lifespan extension isp-1(qm150) elevates ROS and increases HIF-1 target gene expression; loss of hif-1 or aha-1 shortens the extended lifespan of isp-1 mutants, indicating HIF-1 is a key mediator of mitochondrial retrograde longevity signaling. (lee2010inhibitionofrespiration pages 2-3, lee2010inhibitionofrespiration pages 4-4, lee2010inhibitionofrespiration pages 2-2) Lee et al., 2010 (lee2010inhibitionofrespiration pages 2-3, lee2010inhibitionofrespiration pages 4-4, lee2010inhibitionofrespiration pages 2-2)
DAF-16/FOXO Required for full lifespan extension DAF-16 target genes are enriched among transcripts upregulated in isp-1 mutants; DAF-16 shows increased nuclear localization, and daf-16 loss markedly suppresses isp-1 longevity. ROS appears to be an upstream activator, with IMB-2, CST-1/2, BAR-1, and MATH-33 supporting DAF-16-dependent longevity. (senchuk2018activationofdaf16foxo pages 4-6, senchuk2018activationofdaf16foxo pages 9-11, senchuk2018activationofdaf16foxo pages 15-17, senchuk2018activationofdaf16foxo pages 1-2) Senchuk et al., 2018 (senchuk2018activationofdaf16foxo pages 4-6, senchuk2018activationofdaf16foxo pages 9-11, senchuk2018activationofdaf16foxo pages 15-17, senchuk2018activationofdaf16foxo pages 1-2)
SKN-1/Nrf2 Required for lifespan extension SKN-1 target genes are activated in isp-1 mutants, and SKN-1 is required for the increased longevity of mitochondrial mutants including isp-1, supporting a ROS-responsive oxidative stress program downstream of ETC dysfunction. (senchuk2018activationofdaf16foxo pages 17-18) Senchuk et al., 2018 (senchuk2018activationofdaf16foxo pages 17-18)
ATFS-1/mitoUPR Dispensable for adult lifespan extension; required for development in isp-1 background ATFS-1 is necessary for induction of mitoUPR reporters and target genes in isp-1 mutants, but adult-only atfs-1 knockdown does not reduce isp-1 lifespan. In contrast, loss of ATFS-1 during development causes developmental arrest or prevents isp-1 animals from reaching adulthood, indicating a stage-specific requirement. (bennett2014activationofthe pages 1-2, bennett2014activationofthe pages 7-8, wu2018mitochondrialunfoldedprotein pages 2-5, wu2018mitochondrialunfoldedprotein pages 1-2, wu2018mitochondrialunfoldedprotein pages 10-13) Bennett et al., 2014; Wu et al., 2018 (bennett2014activationofthe pages 1-2, bennett2014activationofthe pages 7-8, wu2018mitochondrialunfoldedprotein pages 2-5, wu2018mitochondrialunfoldedprotein pages 1-2, wu2018mitochondrialunfoldedprotein pages 10-13)
Developmental timing (L3/L4 window) Required for establishment of lifespan extension ETC inhibition including isp-1 RNAi extends lifespan only when imposed during larval development, especially by late L3/early L4; similar perturbation in adults fails to produce longevity, indicating a developmentally programmed mitochondrial checkpoint or signaling window. (rea2007relationshipbetweenmitochondrial pages 1-2, rea2007relationshipbetweenmitochondrial pages 7-8, rea2007relationshipbetweenmitochondrial pages 2-3, rea2007relationshipbetweenmitochondrial pages 6-7) Rea et al., 2007 (rea2007relationshipbetweenmitochondrial pages 1-2, rea2007relationshipbetweenmitochondrial pages 7-8, rea2007relationshipbetweenmitochondrial pages 2-3, rea2007relationshipbetweenmitochondrial pages 6-7)

Table: This table summarizes the major signaling pathways and timing requirements linked to isp-1-mediated mitochondrial dysfunction in C. elegans. It distinguishes pathways needed for lifespan extension from those primarily required for development.

6.1. HIF-1 Hypoxia Response Pathway

Lee, Hwang, and Kenyon (2010) demonstrated that isp-1(qm150) mutants exhibit increased expression of HIF-1-dependent target genes, including nhr-57 and F22B5.4 (lee2010inhibitionofrespiration pages 4-4, lee2010inhibitionofrespiration pages 2-2). Loss of hif-1 or RNAi knockdown of aha-1 (HIF-1β) significantly shortened the extended lifespan of isp-1 mutants, establishing HIF-1 as a key mediator of mitochondrial retrograde longevity signaling (lee2010inhibitionofrespiration pages 2-3, lee2010inhibitionofrespiration pages 2-2). Epistasis experiments showed that isp-1 mutations do not further extend lifespan in vhl-1 or egl-9 mutants (which constitutively stabilize HIF-1), consistent with isp-1 acting upstream of or through the HIF-1 pathway (lee2010inhibitionofrespiration pages 2-2).

6.2. DAF-16/FOXO Transcription Factor

Senchuk et al. (2018) found that DAF-16 target genes are significantly enriched among transcripts upregulated in isp-1 mutants, with approximately 50% overlap with genes upregulated in the long-lived insulin/IGF-1 pathway mutant daf-2 (senchuk2018activationofdaf16foxo pages 4-6). DAF-16 shows increased nuclear localization in isp-1 worms, and genetic disruption of daf-16 markedly reduces the lifespan extension (from ~72% to ~19% increase over wild-type) (senchuk2018activationofdaf16foxo pages 9-11). Multiple DAF-16-interacting proteins—including MATH-33, IMB-2 (transportin-1 homolog), CST-1/CST-2 (protein kinases), and BAR-1 (β-catenin homolog)—are required for the full longevity of isp-1 mutants, indicating a complex regulatory network downstream of ROS-mediated DAF-16 activation (senchuk2018activationofdaf16foxo pages 15-17, senchuk2018activationofdaf16foxo pages 1-2).

6.3. SKN-1/Nrf2 Oxidative Stress Response

SKN-1 target genes are also activated in isp-1 mutants, and SKN-1 is required for the increased longevity observed in these animals (senchuk2018activationofdaf16foxo pages 17-18). This indicates that the oxidative stress response mediated by SKN-1/Nrf2 constitutes an additional arm of the mitochondrial retrograde signaling network.

6.4. ATFS-1 and the Mitochondrial Unfolded Protein Response (mitoUPR)

The role of ATFS-1 and the mitoUPR in isp-1 biology is nuanced and somewhat debated. isp-1 mutants exhibit ATFS-1-dependent activation of the mitoUPR, as evidenced by upregulation of the hsp-6p::GFP reporter and endogenous target genes (wu2018mitochondrialunfoldedprotein pages 2-5, bennett2014activationofthe pages 6-6). However, Bennett et al. (2014) demonstrated that knockdown of atfs-1 did not prevent lifespan extension in isp-1(qm150) mutants, leading to the conclusion that the mitoUPR is neither necessary nor sufficient for longevity (bennett2014activationofthe pages 1-2, bennett2014activationofthe pages 7-8). Wu et al. (2018) provided additional resolution by showing that while ATFS-1 is dispensable for adult lifespan maintenance, it is absolutely essential during development—loss of atfs-1 during early development prevents isp-1 worms from reaching adulthood (wu2018mitochondrialunfoldedprotein pages 2-5, wu2018mitochondrialunfoldedprotein pages 1-2). This indicates a stage-specific requirement for mitoUPR activation.

6.5. Developmental Timing Requirement

A critical finding from Rea, Ventura, and Johnson (2007) established that mitochondrial dysfunction-dependent life extension requires perturbation during the L3/L4 larval stage, which coincides with the last somatic cell divisions and massive mitochondrial DNA expansion in C. elegans (rea2007relationshipbetweenmitochondrial pages 1-2, rea2007relationshipbetweenmitochondrial pages 7-8, rea2007relationshipbetweenmitochondrial pages 2-3). When isp-1 RNAi or other ETC inhibition is applied only during adulthood, lifespan is not extended, indicating that a developmental signal initiated during the proliferative larval period is essential for programming the longevity phenotype (rea2007relationshipbetweenmitochondrial pages 6-7).


7. Additional Biological Roles

7.1. Axon Regeneration

Knowlton et al. (2017) identified isp-1 as one of a select subset of electron transport chain genes required for axon regeneration in C. elegans mechanosensory neurons. isp-1(qm150) mutants showed normal axonal development and growth cone formation after laser axotomy but were impaired in subsequent axon extension (knowlton2017aselectsubset pages 1-2). Critically, pan-neuronal overexpression of isp-1 was sufficient to enhance axon regrowth above wild-type levels, indicating that ISP-1-dependent mitochondrial function is rate-limiting for axon regeneration (knowlton2017aselectsubset pages 12-13, knowlton2017aselectsubset pages 9-11). Loss of isp-1 was epistatic to enhanced calcium signaling (egl-19 gain-of-function) and elevated MAP kinase signaling (DLK-1 overexpression), positioning mitochondrial function downstream of or in parallel with these injury response pathways (knowlton2017aselectsubset pages 8-9, knowlton2017aselectsubset pages 7-8).

7.2. Non-Cell Autonomous Signaling from GABAergic Neurons

Recent work by Rathor et al. (2024) demonstrated that isp-1 knockdown specifically in GABAergic neurons extends organismal lifespan and enhances stress resistance through non-cell autonomous mechanisms (rathor2024mitochondrialstressin pages 10-13, rathor2024mitochondrialstressin pages 40-43). DAF-16/FoxO is essential for mediating these systemic effects on lifespan, stress tolerance, mitochondrial homeostasis, and reproductive capacity. The neuropeptide FLP-13, expressed in GABAergic neurons, was identified as a mediator of this non-cell autonomous aging regulation, with isp-1 knockdown and loss of GABA function operating through the same pathway (rathor2024mitochondrialstressin pages 10-13).

7.3. Supercomplex Assembly and Mitochondrial Architecture

ISP-1 serves as a stabilizer of higher-order respiratory supercomplex assemblies (I:III:IV respirasomes). The isp-1(qm150) mutation disrupts supercomplex formation, thereby impairing not only Complex III activity but also complex I function, and potentially increasing ROS production rates (osz2025mutationsofthe pages 19-21).


8. Relationship to Other Mitochondrial Longevity Mutants

isp-1 belongs to a group of C. elegans mitochondrial mutants—alongside clk-1 (ubiquinone biosynthesis), nuo-6 (Complex I subunit), and others—that share the paradoxical property of extending lifespan despite compromised mitochondrial function. Both clk-1 and isp-1 mutants show elevated ROS and require HIF-1 for longevity, though they differ mechanistically: isp-1(qm150) reduces oxygen consumption rate while clk-1(qm30) does not (lee2010inhibitionofrespiration pages 2-3, lee2010inhibitionofrespiration pages 4-4). All three long-lived mitochondrial mutants (clk-1, isp-1, nuo-6) exhibit overlapping transcriptional responses including upregulation of DAF-16 target genes (senchuk2018activationofdaf16foxo pages 1-2).


9. Summary

The isp-1 gene encodes the Rieske iron-sulfur protein, a catalytic subunit of mitochondrial Complex III that is essential for the Q-cycle mechanism of ubiquinol:cytochrome c oxidoreduction. The protein is anchored in the mitochondrial inner membrane by a single transmembrane helix and operates through a "spring-loaded" tether mechanism that enables its 2Fe-2S cluster-containing head domain to shuttle electrons from ubiquinol at the Qo site to cytochrome c1, while simultaneously functioning as a proton-exiting gate. Partial loss of ISP-1 function, as exemplified by the qm150 allele, reduces electron flux through Complex III and triggers a cascade of compensatory mitochondrial retrograde signaling through HIF-1, DAF-16/FOXO, SKN-1/Nrf2, and ATFS-1/mitoUPR pathways—primarily driven by elevated mitochondrial ROS. This signaling, when initiated during a critical developmental window at the L3/L4 larval stage, programs the organism for extended lifespan. Beyond aging, ISP-1 function is rate-limiting for axon regeneration and participates in non-cell autonomous inter-tissue signaling from GABAergic neurons to regulate systemic stress resistance and longevity.

References

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Artifacts

Citations

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  2. osz2025mutationsofthe pages 4-6
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📚 Additional Documentation

Notes

(isp-1-notes.md)

isp-1 (C. elegans) — Gene Review Notes

UniProt: O44512 (O44512_CAEEL) · WormBase: WBGene00002162 / F42G8.12 · NCBI Gene: 177609
Locus: Chromosome IV · Protein: 276 aa · EC 7.1.1.8

Summary / identity

isp-1 encodes the Rieske iron-sulfur protein (ISP), the [2Fe-2S]-cluster-bearing
subunit of mitochondrial respiratory complex III (cytochrome bc1 / ubiquinol–cytochrome
c oxidoreductase). Within complex III it accepts electrons from ubiquinol at the Qo site and
passes them to cytochrome c1, as part of the protonmotive Q-cycle.

  • UniProt names it "Cytochrome b-c1 complex subunit Rieske, mitochondrial"; EC 7.1.1.8;
    belongs to the Rieske iron-sulfur protein family; binds 1 [2Fe-2S] cluster per subunit;
    localized to the mitochondrion inner membrane (isp-1-uniprot.txt, RuleBase RU004494/RU004495).
  • PANTHER family PTHR10134 "CYTOCHROME B-C1 COMPLEX SUBUNIT RIESKE, MITOCHONDRIAL"
    (interpro/panther/PTHR10134/).
  • Independent literature confirmation of identity: "The Caenorhabditis elegans isp-1 gene
    encodes the Rieske iron-sulfur protein subunit of cytochrome c oxidoreductase (complex III of
    the electron transport chain)." PMID:26504246
  • "The nuo-6 and isp-1 genes of C. elegans encode, respectively, subunits of complex I and III
    of the mitochondrial respiratory chain." PMID:21151885

KNOWN (well established)

Core molecular function / localization

  • Rieske [2Fe-2S] iron-sulfur subunit of complex III; electron transfer from ubiquinol to
    cytochrome c1; part of the ubiquinol–cytochrome c reductase reaction (EC 7.1.1.8).
    Cofactor: one [2Fe-2S] cluster (Rieske high-potential type). Source: UniProt
    (isp-1-uniprot.txt: CATALYTIC ACTIVITY, COFACTOR, MISCELLANEOUS "The Rieske protein is a
    high potential 2Fe-2S protein"), Rieske-family conservation (PANTHER PTHR10134, InterPro
    IPR017941 Rieske_2Fe-2S). Reactome R-CEL-611105 (Respiratory electron transport),
    R-CEL-9865881 (Complex III assembly).
  • Mitochondrial inner membrane localization (UniProt SUBCELLULAR LOCATION; single-pass membrane
    anchor + Rieske catalytic head).

Biological process (respiration) — experimentally supported in worm

  • The complex III mutant isp-1(qm150) reduces mitochondrial respiration: it shows impaired
    complex I-dependent (malate) and diminished complex II-dependent (succinate) oxidative
    phosphorylation capacity — expected because complex III is the common downstream acceptor.
    PMID:16920626 and
    PMID:16920626. This is the basis of
    the WormBase IMP annotation to GO:0006122.

The isp-1(qm150) longevity mutant (downstream phenotype, NOT the core MF)

  • isp-1(qm150) is a partial (hypomorphic) loss-of-function missense allele. Phenotypes:
    "low oxygen consumption, decreased sensitivity to ROS, and increased life span."
    PMID:11709184
  • It is one of the classic Mit longevity mutants: "longevity is increased by a partial
    loss-of-function mutation in the mitochondrial complex III subunit gene isp-1."
    PMID:20346072
  • Additional pleiotropic phenotypes of qm150 (developmental rate, pharyngeal pumping, brood
    size, movement, constitutive UPR-mt reporter, CO2 production, OXPHOS, lifespan). PMID:26504246
  • Mechanism of longevity is a mitohormetic superoxide signal, not reduced oxidative damage:
    "the generation of superoxide is elevated in the nuo-6 and isp-1 mitochondrial mutants ...
    this elevation is necessary and sufficient to increase longevity." PMID:21151885 and PMID:21151885
  • qm150 mutation and isp-1(RNAi) act via distinct, separable mechanisms (additive lifespan
    effects). PMID:20346072

NOT KNOWN / knowledge gaps

  • Mechanism linking the Rieske perturbation to the retrograde longevity signal. How the
    specific qm150 substitution in ISP-1 is converted into the elevated-superoxide signal that
    drives the pro-longevity transcriptional program is not molecularly defined. Kaeberlein and
    colleagues localized intragenic suppressors to a conserved six-residue "tether" region and
    proposed a "spring-loaded" gating model, but this is a hypothesis: "The focus on a single
    subunit as causal both in generation and in suppression of diverse pleiotropic phenotypes
    points to a common underlying molecular mechanism, for which we propose a 'spring-loaded'
    model." PMID:26504246. The causal chain from tether-region dynamics
    → altered Qo-site electron transfer/superoxide → downstream gene expression remains open
    (BIOLOGY gap, RESIDUAL_SUBGAP; the textbook complex III function itself is solid).
  • Whether ISP-1 has any function beyond complex III electron transfer (moonlighting) is not
    established; no evidence supports one, and none is claimed here.

GOA annotation review plan (9 annotations)

# Term Evid Action Rationale
1 GO:0016491 oxidoreductase activity IBA MODIFY→GO:0009055 correct but over-general; subunit MF is electron transfer
2 GO:0045275 respiratory chain complex III IBA ACCEPT core CC (complex membership)
3 GO:0006122 mito electron transport ubiquinol→cyt c IBA ACCEPT core BP
4 GO:0005743 mitochondrial inner membrane IEA ACCEPT core CC
5 GO:0008121 quinol-cytochrome-c reductase activity IEA ACCEPT complex-level MF ISP-1 contributes to (core)
6 GO:0016020 membrane IEA MARK_AS_OVER_ANNOTATED uninformative; subsumed by GO:0005743
7 GO:0051537 2 iron, 2 sulfur cluster binding IEA ACCEPT core MF (defining Rieske cofactor)
8 GO:1902600 proton transmembrane transport IEA KEEP_AS_NON_CORE Q-cycle proton translocation is a complex-level consequence, not ISP-1's direct MF
9 GO:0006122 (IMP, PMID:16920626) IMP ACCEPT experimentally supported in worm (complex III respiration defect)

No aging/longevity/behavioral GO annotations are present in the GOA (they are mutant phenotypes,
not curated normal roles), so none need down-weighting there; the longevity biology is captured
in description and knowledge_gaps only.

References verified against PubMed (identity confirmed)

  • PMID:11709184 — Feng, Bussière, Hekimi 2001 Dev Cell 1:633-44 (isp-1(qm150) discovery). VERIFIED.
    NOTE: my first guessed PMID "11740940" for this paper was WRONG (that PMID is a Drosophila
    neuralized paper); corrected to 11709184 via PubMed search + metadata.
  • PMID:21151885 — Yang & Hekimi 2010 PLoS Biol 8:e1000556 (superoxide longevity signal). VERIFIED, full text.
  • PMID:20346072 — Yang & Hekimi 2010 Aging Cell 9:433-47 (two modes; qm150 vs RNAi). VERIFIED.
  • PMID:26504246 — Jafari...Kaeberlein 2015 PNAS 112:E6148-57 (tether suppressors; spring-loaded model). VERIFIED.
  • PMID:16920626 — Falk et al 2006 Curr Biol 16:1641-5 (complex III mutant respiration; IMP source). VERIFIED, full text.

📄 View Raw YAML

id: O44512
gene_symbol: isp-1
product_type: PROTEIN
status: DRAFT
taxon:
  id: NCBITaxon:6239
  label: Caenorhabditis elegans
description: >-
  isp-1 encodes the Rieske iron-sulfur protein (ISP), the [2Fe-2S]-cluster-bearing
  catalytic subunit of mitochondrial respiratory complex III (the cytochrome bc1 /
  ubiquinol-cytochrome c oxidoreductase complex, EC 7.1.1.8). Anchored in the
  mitochondrial inner membrane, its mobile Rieske head domain accepts an electron
  from ubiquinol at the complex III Qo site and delivers it to cytochrome c1,
  performing the electron-transfer step of the protonmotive Q-cycle and thereby
  feeding electrons into the respiratory chain. In C. elegans the partial
  loss-of-function allele isp-1(qm150) is a classic mitochondrial ("Mit") longevity
  mutant: it lowers oxygen consumption and extends lifespan, with the lifespan
  extension driven by an elevated mitochondrial superoxide signal rather than by
  reduced oxidative damage.
references:
  - id: GO_REF:0000002
    title: Gene Ontology annotation through association of InterPro records with GO
      terms
    findings: []
  - id: GO_REF:0000033
    title: Annotation inferences using phylogenetic trees
    findings: []
  - id: GO_REF:0000044
    title: Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location
      vocabulary mapping, accompanied by conservative changes to GO terms applied by
      UniProt
    findings: []
  - id: GO_REF:0000108
    title: Automatic assignment of GO terms using logical inference, based on on inter-ontology
      links
    findings: []
  - id: GO_REF:0000120
    title: Combined Automated Annotation using Multiple IEA Methods
    findings: []
  - id: PMID:16920626
    title: Mitochondrial complex I function modulates volatile anesthetic sensitivity
      in C. elegans.
    findings: []
    reference_review:
      relevance: MEDIUM
      correctness: VERIFIED
      review_notes: >-
        Primarily a complex I / volatile anesthetic study, but includes direct
        respirometry on the complex III mutant isp-1: it shows diminished
        complex II-dependent oxidative phosphorylation, supporting isp-1's role in
        mitochondrial respiration. Source of the WormBase IMP annotation to
        GO:0006122. PMID and content verified against PubMed and cached full text.
  - id: PMID:11709184
    title: Mitochondrial electron transport is a key determinant of life span in Caenorhabditis
      elegans.
    findings: []
    reference_review:
      relevance: HIGH
      correctness: VERIFIED
      review_notes: >-
        Feng, Bussiere & Hekimi 2001 (Dev Cell). Discovery paper for isp-1(qm150):
        identifies the mutation in the C. elegans iron-sulfur protein of complex III
        and establishes low oxygen consumption, decreased ROS sensitivity, and
        increased lifespan. PubMed-verified (note: an initially mis-recollected PMID,
        11740940, is a different Drosophila paper; corrected to 11709184).
  - id: PMID:21151885
    title: A mitochondrial superoxide signal triggers increased longevity in Caenorhabditis
      elegans.
    findings: []
    reference_review:
      relevance: HIGH
      correctness: VERIFIED
      review_notes: >-
        Yang & Hekimi 2010 (PLoS Biol). Establishes that isp-1 (and nuo-6) are
        complex III/I subunits whose partial loss decreases electron transport, and
        that an elevated mitochondrial superoxide signal is necessary and sufficient
        for the longevity. PubMed-verified; full text cached.
  - id: PMID:20346072
    title: Two modes of mitochondrial dysfunction lead independently to lifespan extension
      in Caenorhabditis elegans.
    findings: []
    reference_review:
      relevance: MEDIUM
      correctness: VERIFIED
      review_notes: >-
        Yang & Hekimi 2010 (Aging Cell). Shows the isp-1(qm150) genomic mutation and
        isp-1(RNAi) extend lifespan by distinct, separable mechanisms, and reiterates
        the "partial loss-of-function of a complex III subunit" framing. PubMed-verified.
  - id: PMID:26504246
    title: Tether mutations that restore function and suppress pleiotropic phenotypes
      of the C. elegans isp-1(qm150) Rieske iron-sulfur protein.
    findings: []
    reference_review:
      relevance: HIGH
      correctness: VERIFIED
      review_notes: >-
        Jafari et al. 2015 (PNAS, Kaeberlein lab). Structure-function of ISP-1:
        intragenic suppressors of qm150 map to a conserved six-residue tether region,
        and the authors propose a "spring-loaded" gating model linking a single Rieske
        subunit to pleiotropic phenotypes including longevity. Key source for the
        knowledge gap. PubMed-verified.
  - id: file:worm/isp-1/isp-1-uniprot.txt
    title: UniProtKB O44512 (isp-1, C. elegans) - Cytochrome b-c1 complex subunit Rieske,
      mitochondrial
    findings: []
    reference_review:
      relevance: HIGH
      correctness: VERIFIED
      review_notes: >-
        UniProt entry establishing the Rieske family assignment, EC 7.1.1.8, the
        [2Fe-2S] cofactor (one per subunit), and mitochondrion inner membrane
        localization used to anchor the cofactor and localization annotations.
  - id: file:worm/isp-1/isp-1-deep-research-falcon.md
    title: Falcon (Edison) deep research report for C. elegans isp-1
    findings: []
    reference_review:
      relevance: LOW
      correctness: UNVERIFIED
      review_notes: >-
        Machine-generated deep-research narrative used only as a background pointer;
        no load-bearing claim in this review depends on it. All substantive claims are
        anchored to PubMed-verified primary literature or the UniProt record.
existing_annotations:
  - term:
      id: GO:0016491
      label: oxidoreductase activity
    evidence_type: IBA
    original_reference_id: GO_REF:0000033
    qualifier: enables
    review:
      summary: >-
        Correct but uninformatively general. ISP-1's specific molecular function is
        electron transfer through its Rieske [2Fe-2S] cluster; the complex-level
        oxidoreductase reaction is captured more precisely by GO:0008121.
      action: MODIFY
      reason: >-
        oxidoreductase activity is a high-level parent. The subunit-specific activity
        of the Rieske protein is electron transfer (GO:0009055), enabled via its 2Fe-2S
        cluster; complex III's overall reaction is quinol-cytochrome-c reductase
        activity (GO:0008121), already separately annotated.
      proposed_replacement_terms:
        - id: GO:0009055
          label: electron transfer activity
      propagation_review:
        root_cause: TERM_SCOPING_PROBLEM
        failure_modes:
          - GRANULARITY_MISMATCH
        source_entities:
          - source_id: PANTHER:PTN000015358
            source_label: Rieske / complex III oxidoreductase node
            comment: >-
              IBA propagates the broad parent oxidoreductase activity; the correct
              subunit-level term is electron transfer activity (GO:0009055).
      supported_by:
        - reference_id: PMID:26504246
          supporting_text: >-
            encodes the Rieske iron-sulfur protein subunit of cytochrome c
            oxidoreductase (complex III of the electron transport chain)
  - term:
      id: GO:0045275
      label: respiratory chain complex III
    evidence_type: IBA
    original_reference_id: GO_REF:0000033
    qualifier: part_of
    review:
      summary: >-
        Core annotation. ISP-1 is the Rieske iron-sulfur subunit of respiratory chain
        complex III (cytochrome bc1).
      action: ACCEPT
      reason: >-
        Well established across eukaryotes and in C. elegans; ISP-1 is an integral
        catalytic subunit of complex III.
      supported_by:
        - reference_id: PMID:21151885
          supporting_text: subunits of complex I and III of the mitochondrial respiratory
            chain
        - reference_id: PMID:26504246
          supporting_text: complex III of the electron transport chain
  - term:
      id: GO:0006122
      label: mitochondrial electron transport, ubiquinol to cytochrome c
    evidence_type: IBA
    original_reference_id: GO_REF:0000033
    qualifier: involved_in
    review:
      summary: >-
        Core biological process. Within complex III, ISP-1 transfers electrons from
        ubiquinol to cytochrome c1 (and onward to cytochrome c).
      action: ACCEPT
      reason: >-
        This is the pathway step complex III performs and to which the Rieske subunit
        is central; phylogenetically well supported and consistent with the
        experimental worm data.
      supported_by:
        - reference_id: PMID:21151885
          supporting_text: Partial loss-of-function mutations in these genes decrease
            electron transport
        - reference_id: file:worm/isp-1/isp-1-deep-research-falcon.md
          supporting_text: transfers electrons from ubiquinol to cytochrome c1
  - term:
      id: GO:0005743
      label: mitochondrial inner membrane
    evidence_type: IEA
    original_reference_id: GO_REF:0000044
    qualifier: located_in
    review:
      summary: >-
        Core localization. Complex III, including the Rieske subunit, resides in the
        mitochondrial inner membrane.
      action: ACCEPT
      reason: >-
        UniProt subcellular location; ISP-1 has a single-pass inner-membrane anchor
        with its Rieske head projecting to the intermembrane-space side.
      supported_by:
        - reference_id: file:worm/isp-1/isp-1-uniprot.txt
          supporting_text: "C:mitochondrial inner membrane; IEA:UniProtKB-SubCell"
  - term:
      id: GO:0008121
      label: quinol-cytochrome-c reductase activity
    evidence_type: IEA
    original_reference_id: GO_REF:0000120
    qualifier: enables
    review:
      summary: >-
        Complex III catalytic activity (EC 7.1.1.8) to which ISP-1 contributes the
        essential electron-transfer step. Retained as a core molecular function of the
        subunit in the context of the assembled complex.
      action: ACCEPT
      reason: >-
        ISP-1 is a catalytic subunit of ubiquinol-cytochrome c oxidoreductase; the
        Rieske cluster carries out the electron-transfer half of this reaction. Best
        represented in core_functions as contributes_to (a complex-level activity).
      supported_by:
        - reference_id: file:worm/isp-1/isp-1-uniprot.txt
          supporting_text: EC=7.1.1.8
        - reference_id: PMID:26504246
          supporting_text: subunit of cytochrome c oxidoreductase
  - term:
      id: GO:0016020
      label: membrane
    evidence_type: IEA
    original_reference_id: GO_REF:0000120
    qualifier: located_in
    review:
      summary: >-
        Uninformative general parent, redundant with the specific and correct
        mitochondrial inner membrane annotation (GO:0005743).
      action: MARK_AS_OVER_ANNOTATED
      reason: >-
        "membrane" adds no information beyond the more precise GO:0005743 already
        assigned; it is an over-general IEA byproduct.
  - term:
      id: GO:0051537
      label: 2 iron, 2 sulfur cluster binding
    evidence_type: IEA
    original_reference_id: GO_REF:0000002
    qualifier: enables
    review:
      summary: >-
        Core molecular function. The defining feature of the Rieske protein: it
        coordinates a high-potential [2Fe-2S] cluster, the redox center used for
        electron transfer.
      action: ACCEPT
      reason: >-
        UniProt cofactor annotation and Rieske-family conservation; one [2Fe-2S]
        cluster is bound per subunit.
      supported_by:
        - reference_id: file:worm/isp-1/isp-1-uniprot.txt
          supporting_text: Binds 1 [2Fe-2S] cluster per subunit
        - reference_id: file:worm/isp-1/isp-1-uniprot.txt
          supporting_text: The Rieske protein is a high potential 2Fe-2S protein
  - term:
      id: GO:1902600
      label: proton transmembrane transport
    evidence_type: IEA
    original_reference_id: GO_REF:0000108
    qualifier: involved_in
    review:
      summary: >-
        Complex III couples electron transfer to proton translocation across the inner
        membrane via the Q-cycle. This is a complex-level chemiosmotic outcome, not
        ISP-1's direct molecular function (electron transfer); retained as non-core.
      action: KEEP_AS_NON_CORE
      reason: >-
        Proton translocation is an emergent property of the intact Q-cycle to which
        ISP-1 contributes, rather than an activity ISP-1 performs on its own. Correct
        to retain, but the subunit's core function is electron transfer.
  - term:
      id: GO:0006122
      label: mitochondrial electron transport, ubiquinol to cytochrome c
    evidence_type: IMP
    original_reference_id: PMID:16920626
    qualifier: involved_in
    review:
      summary: >-
        Experimentally supported in C. elegans. The isp-1 complex III mutant shows
        impaired mitochondrial respiration/oxidative phosphorylation, directly
        implicating isp-1 in mitochondrial electron transport.
      action: ACCEPT
      reason: >-
        Falk et al. measured respiration in the isp-1 (complex III) mutant and found
        diminished complex II-dependent oxidative phosphorylation, consistent with a
        defect in the ubiquinol-to-cytochrome c step; core process, experimentally
        anchored.
      supported_by:
        - reference_id: PMID:16920626
          supporting_text_fulltext: in isp-1 (complex III mutant)
          full_text_unavailable: true
        - reference_id: PMID:16920626
          supporting_text_fulltext: somewhat diminished in the complex III ( isp-1 ) mutant
          full_text_unavailable: true
core_functions:
  - description: >-
      Electron-transfer subunit of mitochondrial complex III: within the cytochrome
      bc1 complex, ISP-1's mobile Rieske head accepts an electron from ubiquinol at the
      Qo site and delivers it to cytochrome c1, the electron-transfer step of the
      ubiquinol-cytochrome c reductase (EC 7.1.1.8) reaction in the protonmotive
      Q-cycle.
    supported_by:
      - reference_id: PMID:26504246
        supporting_text: >-
          encodes the Rieske iron-sulfur protein subunit of cytochrome c
          oxidoreductase (complex III of the electron transport chain)
      - reference_id: PMID:21151885
        supporting_text: subunits of complex I and III of the mitochondrial respiratory
          chain
    molecular_function:
      id: GO:0009055
      label: electron transfer activity
    contributes_to_molecular_function:
      id: GO:0008121
      label: quinol-cytochrome-c reductase activity
    directly_involved_in:
      - id: GO:0006122
        label: mitochondrial electron transport, ubiquinol to cytochrome c
    locations:
      - id: GO:0005743
        label: mitochondrial inner membrane
    in_complex:
      id: GO:0045275
      label: respiratory chain complex III
  - description: >-
      Coordinates the Rieske high-potential [2Fe-2S] cluster (one per subunit), the
      redox cofactor that carries out the electron-transfer step of complex III.
    supported_by:
      - reference_id: file:worm/isp-1/isp-1-uniprot.txt
        supporting_text: Binds 1 [2Fe-2S] cluster per subunit
    molecular_function:
      id: GO:0051537
      label: 2 iron, 2 sulfur cluster binding
    in_complex:
      id: GO:0045275
      label: respiratory chain complex III
knowledge_gaps:
  - gap_statement: >-
      The molecular mechanism by which the isp-1(qm150) Rieske substitution is
      converted into the pro-longevity retrograde signal is undetermined. It is
      established that qm150 elevates mitochondrial superoxide and that this elevation
      is necessary and sufficient for the lifespan extension, but the causal chain from
      the specific perturbation of ISP-1 head/tether dynamics, through altered Qo-site
      electron transfer and superoxide production, to the downstream transcriptional
      longevity program has not been resolved.
    boundary: >-
      Firmly established: isp-1 encodes the Rieske [2Fe-2S] subunit of complex III;
      qm150 is a partial loss-of-function allele that lowers oxygen consumption and
      extends lifespan; the longevity requires elevated mitochondrial superoxide
      (abolished by antioxidants, phenocopied by paraquat); and intragenic suppressors
      that restore function map to a conserved six-residue tether region ("spring-loaded"
      model). What is unresolved is the signal-generating step itself.
    gap_kind:
      - BIOLOGY
    dark_aspect: RESIDUAL_SUBGAP
    status: OPEN
    significance: >-
      isp-1(qm150) is a canonical mitohormesis / Mit longevity model. Defining the
      signal-generating step would connect a specific electron-transport-chain lesion to
      a conserved lifespan-extension program with relevance to mitochondrial disease and
      aging.
    resolution: >-
      Structure-guided electron-transfer and superoxide measurements on qm150 and its
      intragenic tether-region suppressors, combined with epistasis to the retrograde
      transcriptional effectors, to identify the step that generates the superoxide
      longevity signal.
    provenance:
      - reference_id: PMID:21151885
        supporting_text: this elevation is necessary and sufficient to increase longevity
      - reference_id: PMID:26504246
        supporting_text: points to a common underlying molecular mechanism, for which
          we propose a