UBE2L6

UniProt ID: G1TUN6
Organism: Oryctolagus cuniculus
Review Status: DRAFT
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Gene Description

UBE2L6 (also known as UBCH8) is an E2 ubiquitin-conjugating enzyme belonging to the UBE2L family. It functions as the principal E2 enzyme in the ISG15 conjugation (ISGylation) pathway, an interferon-stimulated ubiquitin-like modification system that is part of the innate immune defense against viral infection. UBE2L6 accepts activated ISG15 from the E1 enzyme UBA7 and transfers it to target proteins in conjunction with E3 ligases such as HERC5. The protein contains a single UBC catalytic domain with a conserved active-site cysteine that forms a thioester intermediate with ubiquitin or ISG15. While UBE2L6 can also function as an E2 for ubiquitin conjugation in vitro, its primary physiological role is in ISGylation during the type I interferon response.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0005524 ATP binding
IEA
GO_REF:0000104
REMOVE
Summary: ATP binding is not a direct molecular function of E2 ubiquitin-conjugating enzymes. E2 enzymes accept activated ubiquitin (or ubiquitin-like proteins) from E1 activating enzymes via a transthioesterification reaction; it is the E1 enzyme that requires ATP to adenylate ubiquitin. This annotation likely arose from an automated rule transfer that conflates the ATP requirement of the E1 step with the E2 enzyme itself.
Reason: E2 ubiquitin-conjugating enzymes do not bind or hydrolyze ATP. The ATP dependence belongs to the E1 activating enzyme (UBA7/UBE1L for ISG15, or UBA1 for ubiquitin). This is an erroneous IEA annotation.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-uniprot.txt
S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine + [E2 ubiquitin-conjugating enzyme]-L-cysteine = [E1 ubiquitin- activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin- conjugating enzyme]-L-cysteine
GO:0016874 ligase activity
IEA
GO_REF:0000104
REMOVE
Summary: The term 'ligase activity' (GO:0016874) is too broad and arguably misleading for an E2 enzyme. E2 enzymes are transferases (EC 2.3.2.23) that transfer ubiquitin or ubiquitin-like modifiers from an E1 thioester to a substrate. True ligase activity in the ubiquitin system is associated with E3 ubiquitin ligases. The EC number 2.3.2.23 assigned to this protein is in the transferase class, not the ligase class.
Reason: E2 enzymes are classified as transferases (EC 2.3.2.23), not ligases. Calling an E2 a ligase is misleading; the ligase function belongs to E3 enzymes that catalyze the final transfer to substrate lysine residues.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-uniprot.txt
EC=2.3.2.23
GO:0061631 ubiquitin conjugating enzyme activity
IEA
GO_REF:0000003
KEEP AS NON CORE
Summary: UBE2L6 is an E2 ubiquitin-conjugating enzyme with a conserved UBC domain and active-site cysteine. While its primary physiological role is in ISGylation rather than ubiquitin conjugation, UBE2L6 retains E2 enzymatic activity for ubiquitin conjugation and is classified under EC 2.3.2.23. This annotation correctly captures the general enzymatic activity of the protein.
Reason: The ubiquitin conjugating enzyme activity is a valid biochemical activity for this E2 enzyme, but the primary physiological role of UBE2L6 is as an ISG15-specific E2, making this a secondary rather than core function.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-uniprot.txt
Belongs to the ubiquitin-conjugating enzyme family
file:RABIT/G1TUN6/G1TUN6-uniprot.txt
EC=2.3.2.23
GO:0019787 ubiquitin-like protein transferase activity
IEA
GO_REF:0000120
ACCEPT
Summary: UBE2L6 is indeed a ubiquitin-like protein transferase, as it transfers ISG15 (a ubiquitin-like protein) to substrates. This annotation is correct and captures the broader enzymatic category that encompasses both the ubiquitin and ISG15 transferase activities of this E2 enzyme. It is a parent term of the more specific ISG15 transferase activity.
Reason: This correctly captures UBE2L6 as a transferase for ubiquitin-like proteins. Since the more specific ISG15 transferase activity is also annotated, this parent term is redundant but not incorrect. Keeping it as it accurately represents the general enzymatic class.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-uniprot.txt
Ubl conjugation pathway
GO:0032020 ISG15-protein conjugation
IEA
GO_REF:0000107
ACCEPT
Summary: ISG15-protein conjugation (ISGylation) is the core biological process that UBE2L6 participates in. This annotation was transferred from the well-characterized human ortholog (O14933/UBE2L6), which has been experimentally demonstrated to be the dedicated E2 enzyme for ISGylation. The rabbit ortholog, identified by Ensembl Compara, would be expected to retain this conserved function.
Reason: ISGylation is the primary biological process for UBE2L6, supported by ortholog transfer from the experimentally characterized human protein.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-goa.tsv
UniProtKB:O14933|ensembl:ENSP00000287156
GO:0042296 ISG15 transferase activity
IEA
GO_REF:0000120
ACCEPT
Summary: ISG15 transferase activity is the core molecular function of UBE2L6. The human ortholog (O14933) has been experimentally shown to be the dedicated E2 enzyme that transfers ISG15 to substrate proteins in conjunction with E3 ligases. This annotation, transferred via Ensembl Compara and combined automated methods, is well-supported by the conserved UBC domain and orthologous relationship.
Reason: This is the most specific and informative molecular function annotation for UBE2L6, representing its primary enzymatic activity as the ISG15- specific E2 conjugating enzyme.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-uniprot.txt
ISG15 transferase activity; IEA:Ensembl
file:RABIT/G1TUN6/G1TUN6-goa.tsv
UniProtKB:O14933|ensembl:ENSP00000287156|UniProtKB:Q9QZU9|ensembl:ENSMUSP00000099702
GO:0045087 innate immune response
IEA
GO_REF:0000107
KEEP AS NON CORE
Summary: UBE2L6/UBCH8 participates in the innate immune response through its role in ISGylation. ISG15 conjugation is an interferon-stimulated defense mechanism that modifies viral and host proteins as part of the antiviral innate immune response. This annotation, transferred from the human ortholog via Ensembl Compara, correctly places UBE2L6 in the broader biological context of innate immunity.
Reason: While UBE2L6 does participate in the innate immune response through ISGylation, this is a high-level biological process term. The more specific ISG15-protein conjugation term (GO:0032020) more precisely captures the direct biological role. This annotation provides useful biological context but is not the most informative process-level annotation.
Supporting Evidence:
file:RABIT/G1TUN6/G1TUN6-goa.tsv
UniProtKB:O14933|ensembl:ENSP00000287156

Core Functions

UBE2L6 functions as the dedicated E2 conjugating enzyme for ISG15, an interferon-stimulated ubiquitin-like modifier. It accepts activated ISG15 from the E1 enzyme UBA7 via a thioester bond at its active-site cysteine and transfers it to substrate proteins in cooperation with E3 ligases.

Molecular Function:
ISG15 transferase activity
Directly Involved In:

References

Gene Ontology annotation based on Enzyme Commission mapping
Electronic Gene Ontology annotations created by transferring manual GO annotations between related proteins based on shared sequence features
Automatic transfer of experimentally verified manual GO annotation data to orthologs using Ensembl Compara
Combined Automated Annotation using Multiple IEA Methods

Deep Research

Falcon

(G1TUN6-deep-research-falcon.md)
Comprehensive Research Report: Functional Annotation of UBE2L6 (G1TUN6) in Oryctolagus cuniculus Falcon Edison Scientific Literature 30 citations 2 artifacts 2026-06-18T20:00:46.615973

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Comprehensive Research Report: Functional Annotation of UBE2L6 (G1TUN6) in Oryctolagus cuniculus

1. Key Concepts and Definitions: Protein Identity, Family, and Domains

The gene UBE2L6 (UniProt: G1TUN6) from Oryctolagus cuniculus encodes an E2 ubiquitin-conjugating enzyme homologous to the human UBE2L6 (UbcH8), and is definitively established as a member of the ubiquitin-conjugating enzyme family based on sequence, protein family (UBC domain), and domain annotation (Ub_conjugating_enzyme, IPR000608, IPR050113, PF00179). This protein is unambiguously identified in rabbit and matches the expected domain and structural characteristics seen in mammalian orthologs (chen2024elucidatingthemechanism pages 1-3, kang2022thediverserepertoire pages 2-3, yao2026ubiquitinconjugatingenzymesin pages 1-3).

2. Enzymatic Function, Reaction Catalyzed, and Substrate Specificity

UBE2L6 serves as a bifunctional E2 enzyme, central to the ISG15 (interferon-stimulated gene 15) conjugation pathway (ISGylation) and capable of performing ubiquitin conjugation under certain circumstances. Its primary cellular role is in ISGylation, a post-translational modification that covalently links ISG15 to substrate proteins in a three-step enzymatic cascade. UBE2L6 specifically receives ISG15 from the E1 enzyme UBA7 (UBE1L) via a trans-thioesterification reaction. Structural and biochemical evidence show high specificity of UBE2L6 for UBA7 in ISGylation (with lower affinity for the canonical ubiquitin E1 enzyme, UBA1) and for E3 ligases such as HERC5, TRIM25, and HHARI/ARIH1, which confer substrate specificity in the ISGylation system (kang2022thediverserepertoire pages 2-3, tecalcocruz2023proteinisgylationa pages 1-3, mathieu2021herc5andthe pages 2-4).

The catalytic process involves:
- Activation of ISG15 by UBA7 (ATP-dependent),
- Transfer of ISG15 onto UBE2L6 catalytic cysteine through transthiolation,
- Handover to an E3 ligase for covalent (isopeptide bond) attachment to lysine residues on target proteins.

UBE2L6 can also catalyze the ubiquitination of substrate proteins, especially in infection and immune contexts, showing preference (experimentally validated) for K48-linked ubiquitination to promote proteasomal degradation of innate immune sensors. However, in vivo, its ISGylation activity dominates due to cellular pathway specialization (zhu2025porcinereproductiveand pages 2-4, kang2022thediverserepertoire pages 4-6).

Enzymatic Property Molecular Details Citations
Enzyme classification UBE2L6 (also called UbcH8) is an E2 ubiquitin/ubiquitin-like conjugating enzyme of the ubiquitin-conjugating enzyme family. In the ISG15 pathway it functions as the cognate E2 that receives activated ISG15 from the E1 enzyme UBA7/UBE1L; it can also participate in ubiquitination reactions. (kang2022thediverserepertoire pages 2-3, sandy2020morethanmeets pages 1-4, mirzalieva2022isg15andisgylation pages 4-5)
Core catalytic role UBE2L6 acts in the middle of the E1-E2-E3 cascade: activated ISG15 is first adenylated and linked to UBA7, then transferred to the active-site cysteine of UBE2L6, and finally passed to an E3 ligase or directly positioned for ligation to substrate lysines. This is the canonical conjugating-enzyme step of ISGylation. (kang2022thediverserepertoire pages 2-3, tecalcocruz2023proteinisgylationa pages 1-3, mathieu2021herc5andthe pages 2-4)
Catalytic mechanism The mechanism is thiol-based chemistry typical of E2 enzymes: UBA7 forms a thioester with ISG15, UBE2L6 accepts ISG15 by transthiolation/transesterification onto its catalytic cysteine, and downstream transfer yields an isopeptide bond between the C-terminus of ISG15 and a substrate lysine. (tecalcocruz2023proteinisgylationa pages 1-3, mathieu2021herc5andthe pages 2-4, kang2022thediverserepertoire pages 2-3)
E1 enzyme specificity Biochemical and structural work shows that UBE2L6 has much higher affinity for the ISG15 E1 UBA7/UBE1L than for the canonical ubiquitin E1 UBA1, supporting functional specificity for ISGylation in cells even though the enzyme can catalyze ubiquitin transfer in vitro. (sandy2020morethanmeets pages 1-4, kang2022thediverserepertoire pages 2-3, chen2024elucidatingthemechanism pages 1-3)
UBA7-UBE2L6 structural interface Cryo-EM and biochemical studies show that UBA7/UBE1L recruits UBE2L6 through its ubiquitin-fold domain (UFD), while additional contacts involve the SCCH domain and crossover loop. These contacts position the active sites for transfer of activated ISG15 and help explain pathway fidelity. (wallace2023insightsintothe pages 2-3, wallace2023insightsintothe pages 1-2, chen2024elucidatingthemechanism pages 1-3)
Structural basis of specificity Recent structural studies identified determinants that enforce UBA7-UBE2L6 selectivity, including strong UFD-mediated affinity, features of the catalytic cysteine capping loop, and favorable active-site cysteine chemistry. These determinants distinguish UBE2L6 from the closely related UBE2L3. (chen2024elucidatingthemechanism pages 1-3, wallace2023insightsintothe pages 1-2)
Active-site chemistry UBE2L6 contains the catalytic cysteine required for thioester formation with ISG15 (and ubiquitin in ubiquitination settings). Experimental structural trapping used a C86-only UBE2L6 construct, underscoring that this catalytic cysteine is essential for E1-to-E2 transfer. (wallace2023insightsintothe pages 2-3, chen2024elucidatingthemechanism pages 1-3)
E1-E2 reaction intermediate Structural capture of a chemically trapped UBE1L-UBE2L6 complex bound to activated ISG15 revealed how the first transfer steps occur and provided evidence for the transient E1-E2 intermediate that precedes substrate modification. (wallace2023insightsintothe pages 1-2, wallace2023insightsintothe pages 2-3)
Disulfide/thioester-related chemistry In addition to the normal thioester transfer step, UBA7 and UBE2L6 can form a disulfide-linked complex under biochemical conditions; recent work suggests this reflects strong and specific E1-E2 recognition and redox-sensitive control of the ISG15 transfer cascade. (chen2024elucidatingthemechanism pages 1-3)
E3 ligase partners The main E3 ligases functioning with UBE2L6 in ISGylation are HERC5, TRIM25/EFP, and HHARI/ARIH1. HERC5 is broadly active and relatively promiscuous, whereas TRIM25 and HHARI show more substrate selectivity. (kang2022thediverserepertoire pages 2-3, sandy2020morethanmeets pages 1-4, mathieu2021herc5andthe pages 2-4)
HERC5-associated mechanism In the HERC5 pathway, the UBE2L6~ISG15 thioester transfers ISG15 onward for covalent attachment to host or viral substrates. HERC5 can act co-translationally at polysomes, making UBE2L6 part of a ribosome-associated antiviral modification system. (mathieu2021herc5andthe pages 1-2, mathieu2021herc5andthe pages 2-4, tecalcocruz2023proteinisgylationa pages 1-3)
Substrate specificity: ISG15 versus ubiquitin UBE2L6 is best described as bifunctional: it is the principal E2 for ISG15 conjugation but is also capable of ubiquitin conjugation. In cells, its stronger biochemical preference for UBA7 and the interferon-induced ISGylation machinery make ISG15 transfer its dominant defined role. (sandy2020morethanmeets pages 1-4, zhu2025porcinereproductiveand pages 2-4, mirzalieva2022isg15andisgylation pages 4-5)
Substrate attachment chemistry The final product of UBE2L6-driven ISGylation is typically mono-ISGylation at substrate lysines via an isopeptide bond between the ISG15 C-terminal glycine motif and the ฮต-amino group of lysine on the target protein. (tecalcocruz2023proteinisgylationa pages 1-3, mathieu2021herc5andthe pages 2-4)
Ubiquitination capacity Beyond ISGylation, UBE2L6 can function as a ubiquitin-conjugating enzyme and has been implicated in ubiquitin-proteasome-mediated degradation of signaling proteins, especially in infection contexts where it promotes K48-linked ubiquitination of innate immune sensors. (zhu2025porcinereproductiveand pages 2-4, kang2022thediverserepertoire pages 4-6)
Ubiquitin chain linkage types Experimental infection studies indicate that UBE2L6 can promote K48-linked ubiquitination of RIG-I and MDA5, leading to proteasome-dependent degradation. Other literature also supports broader involvement of UBE2L6 in ubiquitin and ISG15 pathway cross-talk, though linkage specificity is best established for K48 in this context. (zhu2025porcinereproductiveand pages 2-4, kang2022thediverserepertoire pages 4-6)
Domain features important for function UBE2L6 is built around the conserved E2/UBC catalytic core. Its function depends on the canonical E2 fold, catalytic cysteine, and precise E1-recognition surfaces that allow discrimination between ISG15 and ubiquitin pathways. (sandy2020morethanmeets pages 1-4, yao2026ubiquitinconjugatingenzymesin pages 1-3, chen2024elucidatingthemechanism pages 1-3)
Pathway fidelity Structural and biochemical analyses show that fidelity in ISG15 signaling depends on selective UBA7-UBE2L6 pairing and recognition of the ISG15 C-terminal ubiquitin-like domain, preventing indiscriminate crossover with the ubiquitin system. (wallace2023insightsintothe pages 1-2, chen2024elucidatingthemechanism pages 1-3, kang2022thediverserepertoire pages 2-3)

Table: This table summarizes the enzymatic properties and molecular mechanism of UBE2L6, emphasizing its role as a dual ubiquitin/ISG15 E2 enzyme and highlighting structural evidence from recent cryo-EM and biochemical studies.

3. Subcellular Localization

UBE2L6 is primarily cytoplasmic, with notable enrichment at sites of active protein translation due to interactions with polysome-associated HERC5, suggesting a role in co-translational ISGylation of nascent proteins. Its distribution can be modulated under stress conditions, including viral infection, and phosphorylation of regulatory motifs affects its subcellular localization (tecalcocruz2023proteinisgylationa pages 1-3, kang2022thediverserepertoire pages 4-6, mathieu2021herc5andthe pages 1-2).

4. Pathways and Biological Rolesโ€”Recent Developments (2023โ€“2025)

UBE2L6 is a key effector of the type I interferon (IFN)โ€“stimulated ISGylation pathway, and plays a crucial role in innate immunity, antiviral defense, inflammation, metabolism, and protein homeostasis. Upon IFN/JAK-STAT pathway activation, UBE2L6 is upregulated alongside ISG15 and E3 ligase partners, underpinning a broad antiviral and cellular stress response.

Mechanisms include:
- Covalent modification of host and viral proteins via ISGylation, suppressing viral replication and propagation.
- UBE2L6-catalyzed ubiquitination of RIG-I/MDA5 (targets of K48-linked ubiquitination and proteasomal degradation) as exploited by specific viruses for immune evasion (e.g., PRRSV, SVA), and modulation of innate sensor stability.
- Promotion of STAT1 ISGylation (in obesity), biasing macrophages toward an M1 inflammatory phenotype.
- Regulation of adipocyte lipid metabolism via ATGL degradation: UBE2L6 ablation in adipose tissue prevents diet-induced obesity and insulin resistance in mice.
- Participation in DNA damage responses and proteostasis, including regulation of newly synthesized and misfolded proteins, partially via cotranslational ISGylation at ribosomes (tecalcocruz2023proteinisgylationa pages 1-3, sandy2020morethanmeets pages 1-4, wei2021adiposespecificknockoutof pages 2-3, mathieu2021herc5andthe pages 2-4).

Pathway/Process Specific Role of UBE2L6 Key Target Proteins/Substrates Biological Outcome
ISGylation pathway and interferon response UBE2L6 is the principal E2 enzyme in the ISG15 conjugation cascade induced by type I interferon; it receives activated ISG15 from UBA7/UBE1L and transfers it to substrates with E3 ligases such as HERC5, TRIM25, and HHARI. In cells, UBE2L6 shows higher functional affinity for the ISG15 pathway than for canonical ubiquitin transfer. (kang2022thediverserepertoire pages 2-3, sandy2020morethanmeets pages 1-4, mathieu2021herc5andthe pages 2-4, mirzalieva2022isg15andisgylation pages 4-5) ISG15; UBA7/UBE1L; HERC5, TRIM25, HHARI; broad host and viral protein substrates Establishes interferon-stimulated ISGylation, a core antiviral and stress-response pathway that remodels protein stability, interactions, and signaling. (kang2022thediverserepertoire pages 2-3, mathieu2021herc5andthe pages 2-4, mirzalieva2022isg15andisgylation pages 4-5)
Innate antiviral immunity UBE2L6 supports antiviral defense by enabling ISGylation of host and viral proteins and by participating in interferon-stimulated effector programs. Its expression is induced with ISG15 pathway genes downstream of IFN/JAK-STAT signaling. (mathieu2021herc5andthe pages 1-2, mathieu2021herc5andthe pages 2-4, kang2022thediverserepertoire pages 4-6) Viral proteins such as influenza NP and other newly synthesized viral proteins via HERC5-associated cotranslational ISGylation; host antiviral signaling proteins Restricts viral protein synthesis, assembly, and replication, and amplifies innate immune responsiveness. In some virus-host contexts, pathogens can hijack UBE2L6 to weaken immunity. (mathieu2021herc5andthe pages 2-4, kang2022thediverserepertoire pages 4-6, zhu2025porcinereproductiveand pages 2-4)
Regulation of RIG-I/MDA5 signaling UBE2L6 modulates RIG-I-like receptor signaling through ubiquitin/ISG15-dependent mechanisms. During PRRSV infection, elevated UBE2L6 promotes K48-linked ubiquitination and proteasomal loss of RIG-I and MDA5, aided by viral NSP5; broader ISG15 literature also links ISGylation to RIG-I/MDA5 regulation. (zhu2025porcinereproductiveand pages 2-4, kang2022thediverserepertoire pages 4-6) RIG-I, MDA5, PRRSV NSP5, K48-linked ubiquitin, ISG15 Suppresses type I IFN and ISG expression during PRRSV infection, facilitating viral replication; demonstrates that UBE2L6 can either support host defense or be co-opted by viruses. (zhu2025porcinereproductiveand pages 2-4)
STAT1 signaling and macrophage polarization In obese mouse models, Ube2L6 promotes STAT1 ISGylation/ISG15-dependent activation, increasing STAT1 abundance and phosphorylation and biasing macrophages toward a pro-inflammatory M1 state. (li2024ube2l6promotesm1 pages 2-3, li2024ube2l6promotesm1 pages 3-4) STAT1, ISG15, macrophage polarization machinery Enhances M1 macrophage polarization, inflammatory cytokine production, and obesity-associated inflammation; Ube2L6 deficiency shifts macrophages away from the M1 phenotype. (li2024ube2l6promotesm1 pages 2-3, li2024ube2l6promotesm1 pages 3-4)
Adipocyte metabolism and lipid regulation In adipose tissue, Ube2L6 negatively regulates ATGL stability and thereby restrains lipolysis. Adipose-specific Ube2l6 knockout in mice increases ATGL stability and alters adipocyte size and differentiation under high-fat diet conditions. Human obesity datasets/tissues also show inverse UBE2L6-ATGL association and positive correlation with BMI. (wei2021adiposespecificknockoutof pages 2-3) ATGL/PNPLA2; adipocyte lipid-droplet/lipolysis machinery Promotes diet-induced obesity, insulin resistance, hepatic steatosis, and adipose expansion when elevated; loss of Ube2L6 improves metabolic phenotype in mice. (wei2021adiposespecificknockoutof pages 2-3)
Protein quality control and proteostasis Through the ISG15 system, UBE2L6 contributes to proteostasis by promoting cotranslational ISGylation of newly synthesized proteins together with HERC5, altering ubiquitin-dependent turnover and helping manage misfolded, viral, or stress-induced substrates. Reduced UBE2L6 lowers ISGylation and can increase ubiquitination in some cancer-cell models. (tecalcocruz2023proteinisgylationa pages 1-3, kang2022thediverserepertoire pages 4-6) Newly synthesized proteins, ubiquitin, ISG15, HERC5-associated polysome substrates, Ubc13 and other proteostasis-linked factors Regulates stability and turnover of protein substrates, restricts nascent viral proteins, and contributes to the balance between ISGylation and ubiquitin-proteasome pathways. (tecalcocruz2023proteinisgylationa pages 1-3, kang2022thediverserepertoire pages 4-6)
DNA damage response UBE2L6 participates in the ISG15 system increasingly linked to genome stability. Reviews identify UBE2L6-dependent ISGylation as part of pathways affecting p53 signaling, replication stress responses, and DNA repair-associated proteome remodeling. (sandy2020morethanmeets pages 1-4, kang2022thediverserepertoire pages 4-6) ISG15 pathway components; DDR-associated proteins including p53-network and replication/repair factors discussed in ISG15 studies Suggests a role in maintaining genome stability and modulating DNA damage responses, though many UBE2L6-specific substrates in this context remain incompletely defined. (sandy2020morethanmeets pages 1-4, kang2022thediverserepertoire pages 4-6)
Autophagy regulation UBE2L6 contributes indirectly to autophagy-related control through ISGylation-dependent regulation of proteins such as BECN1 and by modulating antiviral signaling nodes that connect to autophagic degradation pathways. ISG15 literature also links ISGylation to RIG-I-associated autophagy control. (tecalcocruz2023proteinisgylationa pages 1-3, kang2022thediverserepertoire pages 4-6) BECN1, RIG-I-associated complexes, ISG15-modified stress-response proteins Can influence antiviral autophagy and selective degradation pathways, thereby integrating innate immune signaling with cellular quality-control responses. (tecalcocruz2023proteinisgylationa pages 1-3, kang2022thediverserepertoire pages 4-6)

Table: This table summarizes experimentally supported and review-backed roles of UBE2L6 across interferon signaling, antiviral immunity, metabolism, inflammation, and proteostasis. It is useful for functional annotation because it links pathway membership to specific substrates and biological outcomes.

5. Up-to-Date Applications and Real-World Implications (2023โ€“2024)

Recent in vivo mouse studies demonstrate that:
- Adipose-specific knockout of Ube2l6 reduces obesity and insulin resistance via increased ATGL stability (wei2021adiposespecificknockoutof pages 2-3).
- Ube2l6 knockdown modulates macrophage polarization and inflammation in metabolic disease settings (li2024ube2l6promotesm1 pages 2-3).
- UBE2L6 is upregulated by type I IFN responses and in metabolic and cardiovascular disease risk tissue (kang2022thediverserepertoire pages 4-6).

6. Expert Consensus and Authoritative Reviews

  • Structural (cryo-EM) and mechanistic studies in 2023โ€“2024 (Wallace et al., Nature Commun. 2023; Chen et al., bioRxiv 2024) define UBA7-UBE2L6-ISG15 specificity and delineate the molecular determinants of ISGylation versus ubiquitination (wallace2023insightsintothe pages 1-2, chen2024elucidatingthemechanism pages 1-3).
  • Functional annotation reviews agree on major roles in ISGylation, innate immunity, viral restriction, and metabolic regulation (kang2022thediverserepertoire pages 2-3, mathieu2021herc5andthe pages 1-2).

7. Relevant Statistics and Data from Recent Studies

  • Adipose tissue Ube2l6 RNA and protein expression are significantly increased in obese mice and humans, exhibiting strong negative correlation with ATGL protein levels and positive correlation with BMI (wei2021adiposespecificknockoutof pages 2-3).
  • Macrophage polarization and STAT1 activation are significantly reduced in Ube2l6-deficient mouse models, reducing inflammation in high-fat dietโ€“induced obesity (li2024ube2l6promotesm1 pages 2-3).
  • UBE2L6 expression is robustly induced by type I IFN and viral infection in multiple cell types (kang2022thediverserepertoire pages 2-3, mathieu2021herc5andthe pages 1-2).

8. URLs and Publication References

  • Wallace et al., Nature Commun. 2023. https://doi.org/10.1038/s41467-023-43711-3
  • Chen et al., bioRxiv 2024. https://doi.org/10.1101/2024.11.07.622398
  • Mathieu et al., Viruses 2021. https://doi.org/10.3390/v13061102
  • Kang et al., Exp Mol Med 2022. https://doi.org/10.1038/s12276-022-00872-3
  • Tecalco-Cruz et al., Explor Target Antitumor Ther 2023. https://doi.org/10.37349/etat.2023.00162
  • Wei et al., J Pharmacol Sci 2021. https://doi.org/10.1016/j.jphs.2020.12.008
  • Li et al., Obes Facts 2024. https://doi.org/10.1159/000533966
  • Zhu et al., Vet Res 2025. https://doi.org/10.1186/s13567-025-01558-0
  • Gonzรกlez-Amor et al., Front Cell Dev Biol 2023. https://doi.org/10.3389/fcell.2023.1128594

This comprehensive annotation integrates recent advances (up to 2025), with authoritative structural, biochemical, and physiological evidence contextualized for O. cuniculus UBE2L6. Current literature and data demonstrate its essential cellular roles in ISGylation, metabolism, and antiviral immunity, all underpinned by precise enzymatic mechanisms and pathway integration. Further details and primary data can be found in the referenced publications and database entries (see URLs above).

References

  1. (chen2024elucidatingthemechanism pages 1-3): Pei-Tzu Chen, Jia-Yin Yeh, Jui-Hsia Weng, and Kuen-Phon Wu. Elucidating the mechanism underlying uba7โ€ขube2l6 disulfide complex formation. bioRxiv, Nov 2024. URL: https://doi.org/10.1101/2024.11.07.622398, doi:10.1101/2024.11.07.622398. This article has 1 citations.

  2. (kang2022thediverserepertoire pages 2-3): Ji An Kang, Yoon Jung Kim, and Young Joo Jeon. The diverse repertoire of isg15: more intricate than initially thought. Experimental & Molecular Medicine, 54:1779-1792, Nov 2022. URL: https://doi.org/10.1038/s12276-022-00872-3, doi:10.1038/s12276-022-00872-3. This article has 108 citations and is from a peer-reviewed journal.

  3. (yao2026ubiquitinconjugatingenzymesin pages 1-3): Zhiyang Yao, Ti Peng, Hao Dong, Yong Liao, Kai Miao, Jiang-Jiang Qin, and Xiaoqing Guan. Ubiquitin-conjugating enzymes in cancer. International Journal of Biological Sciences, 22:3244-3271, Mar 2026. URL: https://doi.org/10.7150/ijbs.130297, doi:10.7150/ijbs.130297. This article has 46 citations and is from a peer-reviewed journal.

  4. (tecalcocruz2023proteinisgylationa pages 1-3): Angeles C. Tecalco-Cruz and Jesรบs Zepeda-Cervantes. Protein isgylation: a posttranslational modification with implications for malignant neoplasms. Exploration of Targeted Anti-tumor Therapy, 4:699-715, Aug 2023. URL: https://doi.org/10.37349/etat.2023.00162, doi:10.37349/etat.2023.00162. This article has 14 citations.

  5. (mathieu2021herc5andthe pages 2-4): Nicholas A. Mathieu, Ermela Paparisto, Stephen D. Barr, and Donald E. Spratt. Herc5 and the isgylation pathway: critical modulators of the antiviral immune response. Viruses, 13:1102, Jun 2021. URL: https://doi.org/10.3390/v13061102, doi:10.3390/v13061102. This article has 91 citations.

  6. (zhu2025porcinereproductiveand pages 2-4): Zhenbang Zhu, Lulu Chen, Meng Zhang, Qianwen Lin, Yifan Yan, Wenqiang Wang, Wei Wen, Zhendong Zhang, and Xiangdong Li. Porcine reproductive and respiratory syndrome virus nsp5 exploited ube2l6 to promote viral replication via antagonising host rlrs and isgylation. Veterinary Research, Jul 2025. URL: https://doi.org/10.1186/s13567-025-01558-0, doi:10.1186/s13567-025-01558-0. This article has 5 citations and is from a highest quality peer-reviewed journal.

  7. (kang2022thediverserepertoire pages 4-6): Ji An Kang, Yoon Jung Kim, and Young Joo Jeon. The diverse repertoire of isg15: more intricate than initially thought. Experimental & Molecular Medicine, 54:1779-1792, Nov 2022. URL: https://doi.org/10.1038/s12276-022-00872-3, doi:10.1038/s12276-022-00872-3. This article has 108 citations and is from a peer-reviewed journal.

  8. (sandy2020morethanmeets pages 1-4): Zac Sandy, Isabelle Cristine da Costa, and Christine K. Schmidt. More than meets the isg15: emerging roles in the dna damage response and beyond. Biomolecules, 10:1557, Nov 2020. URL: https://doi.org/10.3390/biom10111557, doi:10.3390/biom10111557. This article has 65 citations.

  9. (mirzalieva2022isg15andisgylation pages 4-5): Oygul Mirzalieva, Meredith Juncker, Joshua Schwartzenburg, and Shyamal Desai. Isg15 and isgylation in human diseases. Cells, 11:538, Feb 2022. URL: https://doi.org/10.3390/cells11030538, doi:10.3390/cells11030538. This article has 130 citations.

  10. (wallace2023insightsintothe pages 2-3): Iona Wallace, Kheewoong Baek, J. Rajan Prabu, Ronnald Vollrath, Susanne von Gronau, Brenda A. Schulman, and Kirby N. Swatek. Insights into the isg15 transfer cascade by the ube1l activating enzyme. Nature Communications, Dec 2023. URL: https://doi.org/10.1038/s41467-023-43711-3, doi:10.1038/s41467-023-43711-3. This article has 31 citations and is from a highest quality peer-reviewed journal.

  11. (wallace2023insightsintothe pages 1-2): Iona Wallace, Kheewoong Baek, J. Rajan Prabu, Ronnald Vollrath, Susanne von Gronau, Brenda A. Schulman, and Kirby N. Swatek. Insights into the isg15 transfer cascade by the ube1l activating enzyme. Nature Communications, Dec 2023. URL: https://doi.org/10.1038/s41467-023-43711-3, doi:10.1038/s41467-023-43711-3. This article has 31 citations and is from a highest quality peer-reviewed journal.

  12. (mathieu2021herc5andthe pages 1-2): Nicholas A. Mathieu, Ermela Paparisto, Stephen D. Barr, and Donald E. Spratt. Herc5 and the isgylation pathway: critical modulators of the antiviral immune response. Viruses, 13:1102, Jun 2021. URL: https://doi.org/10.3390/v13061102, doi:10.3390/v13061102. This article has 91 citations.

  13. (wei2021adiposespecificknockoutof pages 2-3): Weiping Wei, Yunqian Li, Yongyong Li, and Daoyuan Li. Adipose-specific knockout of ubiquitin-conjugating enzyme e2l6 (ube2l6) reduces diet-induced obesity, insulin resistance, and hepatic steatosis. Journal of pharmacological sciences, 145 4:327-334, Apr 2021. URL: https://doi.org/10.1016/j.jphs.2020.12.008, doi:10.1016/j.jphs.2020.12.008. This article has 18 citations and is from a peer-reviewed journal.

  14. (li2024ube2l6promotesm1 pages 2-3): Yunqian Li, Xiao Dong, Wenqian He, Huibiao Quan, Kaining Chen, Chaoping Cen, and Weiping Wei. Ube2l6 promotes m1 macrophage polarization in high-fat diet-fed obese mice via isgylation of stat1 to trigger stat1 activation. Obesity Facts, 17:24-36, Oct 2024. URL: https://doi.org/10.1159/000533966, doi:10.1159/000533966. This article has 17 citations and is from a peer-reviewed journal.

  15. (li2024ube2l6promotesm1 pages 3-4): Yunqian Li, Xiao Dong, Wenqian He, Huibiao Quan, Kaining Chen, Chaoping Cen, and Weiping Wei. Ube2l6 promotes m1 macrophage polarization in high-fat diet-fed obese mice via isgylation of stat1 to trigger stat1 activation. Obesity Facts, 17:24-36, Oct 2024. URL: https://doi.org/10.1159/000533966, doi:10.1159/000533966. This article has 17 citations and is from a peer-reviewed journal.

Artifacts

Citations

  1. chen2024elucidatingthemechanism pages 1-3
  2. zhu2025porcinereproductiveand pages 2-4
  3. wei2021adiposespecificknockoutof pages 2-3
  4. kang2022thediverserepertoire pages 4-6
  5. kang2022thediverserepertoire pages 2-3
  6. yao2026ubiquitinconjugatingenzymesin pages 1-3
  7. tecalcocruz2023proteinisgylationa pages 1-3
  8. sandy2020morethanmeets pages 1-4
  9. wallace2023insightsintothe pages 2-3
  10. wallace2023insightsintothe pages 1-2
  11. https://doi.org/10.1038/s41467-023-43711-3
  12. https://doi.org/10.1101/2024.11.07.622398
  13. https://doi.org/10.3390/v13061102
  14. https://doi.org/10.1038/s12276-022-00872-3
  15. https://doi.org/10.37349/etat.2023.00162
  16. https://doi.org/10.1016/j.jphs.2020.12.008
  17. https://doi.org/10.1159/000533966
  18. https://doi.org/10.1186/s13567-025-01558-0
  19. https://doi.org/10.3389/fcell.2023.1128594
  20. https://doi.org/10.1101/2024.11.07.622398,
  21. https://doi.org/10.1038/s12276-022-00872-3,
  22. https://doi.org/10.7150/ijbs.130297,
  23. https://doi.org/10.37349/etat.2023.00162,
  24. https://doi.org/10.3390/v13061102,
  25. https://doi.org/10.1186/s13567-025-01558-0,
  26. https://doi.org/10.3390/biom10111557,
  27. https://doi.org/10.3390/cells11030538,
  28. https://doi.org/10.1038/s41467-023-43711-3,
  29. https://doi.org/10.1016/j.jphs.2020.12.008,
  30. https://doi.org/10.1159/000533966,

๐Ÿ“„ View Raw YAML

id: G1TUN6
gene_symbol: UBE2L6
product_type: PROTEIN
status: DRAFT
taxon:
  id: NCBITaxon:9986
  label: Oryctolagus cuniculus
description: >-
  UBE2L6 (also known as UBCH8) is an E2 ubiquitin-conjugating enzyme belonging
  to the UBE2L family. It functions as the principal E2 enzyme in the ISG15
  conjugation (ISGylation) pathway, an interferon-stimulated ubiquitin-like
  modification system that is part of the innate immune defense against viral
  infection. UBE2L6 accepts activated ISG15 from the E1 enzyme UBA7 and
  transfers it to target proteins in conjunction with E3 ligases such as HERC5.
  The protein contains a single UBC catalytic domain with a conserved active-site
  cysteine that forms a thioester intermediate with ubiquitin or ISG15. While
  UBE2L6 can also function as an E2 for ubiquitin conjugation in vitro, its
  primary physiological role is in ISGylation during the type I interferon
  response.
existing_annotations:
- term:
    id: GO:0005524
    label: ATP binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000104
  qualifier: enables
  review:
    summary: >-
      ATP binding is not a direct molecular function of E2 ubiquitin-conjugating
      enzymes. E2 enzymes accept activated ubiquitin (or ubiquitin-like
      proteins) from E1 activating enzymes via a transthioesterification
      reaction; it is the E1 enzyme that requires ATP to adenylate ubiquitin.
      This annotation likely arose from an automated rule transfer that
      conflates the ATP requirement of the E1 step with the E2 enzyme itself.
    action: REMOVE
    reason: >-
      E2 ubiquitin-conjugating enzymes do not bind or hydrolyze ATP. The ATP
      dependence belongs to the E1 activating enzyme (UBA7/UBE1L for ISG15, or
      UBA1 for ubiquitin). This is an erroneous IEA annotation.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-uniprot.txt
      supporting_text: >-
        S-ubiquitinyl-[E1 ubiquitin-activating enzyme]-L-cysteine +
        [E2 ubiquitin-conjugating enzyme]-L-cysteine = [E1 ubiquitin-
        activating enzyme]-L-cysteine + S-ubiquitinyl-[E2 ubiquitin-
        conjugating enzyme]-L-cysteine
- term:
    id: GO:0016874
    label: ligase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000104
  qualifier: enables
  review:
    summary: >-
      The term 'ligase activity' (GO:0016874) is too broad and arguably
      misleading for an E2 enzyme. E2 enzymes are transferases (EC 2.3.2.23)
      that transfer ubiquitin or ubiquitin-like modifiers from an E1 thioester
      to a substrate. True ligase activity in the ubiquitin system is associated
      with E3 ubiquitin ligases. The EC number 2.3.2.23 assigned to this
      protein is in the transferase class, not the ligase class.
    action: REMOVE
    reason: >-
      E2 enzymes are classified as transferases (EC 2.3.2.23), not ligases.
      Calling an E2 a ligase is misleading; the ligase function belongs to E3
      enzymes that catalyze the final transfer to substrate lysine residues.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-uniprot.txt
      supporting_text: "EC=2.3.2.23"
- term:
    id: GO:0061631
    label: ubiquitin conjugating enzyme activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000003
  qualifier: enables
  review:
    summary: >-
      UBE2L6 is an E2 ubiquitin-conjugating enzyme with a conserved UBC domain
      and active-site cysteine. While its primary physiological role is in
      ISGylation rather than ubiquitin conjugation, UBE2L6 retains E2 enzymatic
      activity for ubiquitin conjugation and is classified under EC 2.3.2.23.
      This annotation correctly captures the general enzymatic activity of the
      protein.
    action: KEEP_AS_NON_CORE
    reason: >-
      The ubiquitin conjugating enzyme activity is a valid biochemical activity
      for this E2 enzyme, but the primary physiological role of UBE2L6 is as an
      ISG15-specific E2, making this a secondary rather than core function.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-uniprot.txt
      supporting_text: "Belongs to the ubiquitin-conjugating enzyme family"
    - reference_id: file:RABIT/G1TUN6/G1TUN6-uniprot.txt
      supporting_text: "EC=2.3.2.23"
- term:
    id: GO:0019787
    label: ubiquitin-like protein transferase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: >-
      UBE2L6 is indeed a ubiquitin-like protein transferase, as it transfers
      ISG15 (a ubiquitin-like protein) to substrates. This annotation is
      correct and captures the broader enzymatic category that encompasses both
      the ubiquitin and ISG15 transferase activities of this E2 enzyme. It is
      a parent term of the more specific ISG15 transferase activity.
    action: ACCEPT
    reason: >-
      This correctly captures UBE2L6 as a transferase for ubiquitin-like
      proteins. Since the more specific ISG15 transferase activity is also
      annotated, this parent term is redundant but not incorrect. Keeping it
      as it accurately represents the general enzymatic class.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-uniprot.txt
      supporting_text: "Ubl conjugation pathway"
- term:
    id: GO:0032020
    label: ISG15-protein conjugation
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: involved_in
  review:
    summary: >-
      ISG15-protein conjugation (ISGylation) is the core biological process
      that UBE2L6 participates in. This annotation was transferred from the
      well-characterized human ortholog (O14933/UBE2L6), which has been
      experimentally demonstrated to be the dedicated E2 enzyme for ISGylation.
      The rabbit ortholog, identified by Ensembl Compara, would be expected to
      retain this conserved function.
    action: ACCEPT
    reason: >-
      ISGylation is the primary biological process for UBE2L6, supported by
      ortholog transfer from the experimentally characterized human protein.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-goa.tsv
      supporting_text: "UniProtKB:O14933|ensembl:ENSP00000287156"
- term:
    id: GO:0042296
    label: ISG15 transferase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: >-
      ISG15 transferase activity is the core molecular function of UBE2L6.
      The human ortholog (O14933) has been experimentally shown to be the
      dedicated E2 enzyme that transfers ISG15 to substrate proteins in
      conjunction with E3 ligases. This annotation, transferred via Ensembl
      Compara and combined automated methods, is well-supported by the
      conserved UBC domain and orthologous relationship.
    action: ACCEPT
    reason: >-
      This is the most specific and informative molecular function annotation
      for UBE2L6, representing its primary enzymatic activity as the ISG15-
      specific E2 conjugating enzyme.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-uniprot.txt
      supporting_text: "ISG15 transferase activity; IEA:Ensembl"
    - reference_id: file:RABIT/G1TUN6/G1TUN6-goa.tsv
      supporting_text: "UniProtKB:O14933|ensembl:ENSP00000287156|UniProtKB:Q9QZU9|ensembl:ENSMUSP00000099702"
- term:
    id: GO:0045087
    label: innate immune response
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: involved_in
  review:
    summary: >-
      UBE2L6/UBCH8 participates in the innate immune response through its role
      in ISGylation. ISG15 conjugation is an interferon-stimulated defense
      mechanism that modifies viral and host proteins as part of the antiviral
      innate immune response. This annotation, transferred from the human
      ortholog via Ensembl Compara, correctly places UBE2L6 in the broader
      biological context of innate immunity.
    action: KEEP_AS_NON_CORE
    reason: >-
      While UBE2L6 does participate in the innate immune response through
      ISGylation, this is a high-level biological process term. The more
      specific ISG15-protein conjugation term (GO:0032020) more precisely
      captures the direct biological role. This annotation provides useful
      biological context but is not the most informative process-level
      annotation.
    supported_by:
    - reference_id: file:RABIT/G1TUN6/G1TUN6-goa.tsv
      supporting_text: "UniProtKB:O14933|ensembl:ENSP00000287156"
core_functions:
- description: >-
    UBE2L6 functions as the dedicated E2 conjugating enzyme for ISG15, an
    interferon-stimulated ubiquitin-like modifier. It accepts activated ISG15
    from the E1 enzyme UBA7 via a thioester bond at its active-site cysteine
    and transfers it to substrate proteins in cooperation with E3 ligases.
  molecular_function:
    id: GO:0042296
    label: ISG15 transferase activity
  directly_involved_in:
  - id: GO:0032020
    label: ISG15-protein conjugation
references:
- id: GO_REF:0000003
  title: Gene Ontology annotation based on Enzyme Commission mapping
  findings: []
- id: GO_REF:0000104
  title: Electronic Gene Ontology annotations created by transferring manual GO annotations
    between related proteins based on shared sequence features
  findings: []
- id: GO_REF:0000107
  title: Automatic transfer of experimentally verified manual GO annotation data to
    orthologs using Ensembl Compara
  findings: []
- id: GO_REF:0000120
  title: Combined Automated Annotation using Multiple IEA Methods
  findings: []