RIC7

UniProt ID: F4JLB7
Organism: Arabidopsis thaliana
Review Status: COMPLETE
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Gene Description

RIC7 (ROP-interactive CRIB motif-containing protein 7; locus At4g28560; UniProt F4JLB7) is a member of the 11-protein RIC family in Arabidopsis thaliana. RIC proteins are plant-specific effectors of ROP (Rho of Plants) GTPases, defined by a conserved CRIB (Cdc42/Rac-interactive binding) domain that mediates preferential binding to the GTP-bound (active) form of ROP proteins. RIC7 also contains leucine-rich repeat (LRR) domains, placing it in the receptor-like protein (RLP) family. RIC7 functions as a signaling adaptor downstream of ROP2, negatively regulating both light-induced stomatal opening and ABA-induced stomatal closure in guard cells. Upon light-stimulated ROP2 activation, RIC7 translocates from the nucleus to the plasma membrane, where it directly binds and inhibits the exocyst subunit Exo70B1, thereby restraining vesicle trafficking required for rapid stomatal aperture changes. The ROP2-RIC7-Exo70B1 signaling module provides a brake mechanism that fine-tunes stomatal kinetics to balance CO2 uptake against water loss. RIC7 is highly expressed in guard cells and has also been implicated in pollen tube growth through its interaction with ROP GTPases.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0005886 plasma membrane
IBA
GO_REF:0000033
ACCEPT
Summary: RIC7 dynamically localizes to the plasma membrane region in guard cells upon light stimulation or co-expression with constitutively active ROP2 (Hong et al. 2016). The IBA annotation from phylogenetic inference is consistent with the experimentally demonstrated plasma membrane localization in guard cells.
Reason: Plasma membrane localization of RIC7 is well supported by confocal microscopy of fluorescently tagged RIC7 in Vicia faba guard cells, showing light-dependent and ROP2-activity-dependent relocalization to the plasma membrane. The phylogenetic inference is consistent with experimental data.
GO:0038023 signaling receptor activity
IBA
GO_REF:0000033
MARK AS OVER ANNOTATED
Summary: RIC7 is not a signaling receptor per se. It is an intracellular signaling adaptor/effector that binds activated ROP GTPases via its CRIB domain and transduces their signals to downstream targets. While it contains LRR domains typical of receptor-like proteins, there is no evidence that RIC7 itself perceives extracellular ligands or has signaling receptor activity. The IBA annotation likely propagated from receptor-like protein family members that do have receptor activity, but RIC7 lacks a kinase domain and functions as a cytoplasmic effector rather than a receptor.
Reason: The phylogenetic annotation appears to have propagated receptor activity from LRR receptor-like kinase or receptor-like protein family members. However, RIC7 is functionally characterized as an intracellular ROP GTPase effector, not a signaling receptor. Its CRIB motif mediates intracellular protein-protein interactions with activated ROP2, not extracellular ligand perception. Small GTPase binding (GO:0031267) would be more appropriate for this protein.
GO:0016020 membrane
IEA
GO_REF:0000044
KEEP AS NON CORE
Summary: UniProt computationally predicts F4JLB7 as a single-pass type I membrane protein based on ARBA analysis. The protein does contain a predicted signal peptide and transmembrane domain. Experimental evidence supports plasma membrane localization in guard cells, so the general membrane annotation is consistent but less informative than the more specific plasma membrane annotation.
Reason: The annotation is correct but overly broad. The protein localizes to the plasma membrane specifically, and more specific annotations exist (GO:0005886, GO:0016324). This IEA annotation is subsumed by those.
GO:0016020 membrane
TAS
PMID:12068095
Prediction of glycosylphosphatidylinositol-anchored proteins...
KEEP AS NON CORE
Summary: PMID:12068095 (Borner et al. 2002) is a genomic analysis predicting GPI-anchored proteins in Arabidopsis. The paper identified RIC7 (At4g28560) as a predicted plasma membrane receptor-like protein. The membrane localization is consistent with this computational prediction and with subsequent experimental evidence.
Reason: The annotation is broadly correct but nonspecific. More informative localization terms (plasma membrane, apical plasma membrane) are available from other annotations. The TAS evidence is based on computational predictions of GPI-anchoring and membrane association rather than direct experimental demonstration.
GO:0016020 membrane
TAS
PMID:12805588
Identification of glycosylphosphatidylinositol-anchored prot...
KEEP AS NON CORE
Summary: PMID:12805588 (Borner et al. 2003) is a proteomic and genomic analysis of GPI-anchored proteins in Arabidopsis that identified receptor-like proteins including those at the At4g28560 locus. The membrane annotation is consistent with these findings.
Reason: Same rationale as the other membrane annotations. The annotation is correct but subsumed by more specific plasma membrane and apical plasma membrane annotations from direct experimental evidence.
GO:0009507 chloroplast
ISM
GO_REF:0000122
MARK AS OVER ANNOTATED
Summary: The chloroplast localization is based on AtSubP computational prediction (ISM evidence). There is no experimental evidence supporting chloroplast localization for RIC7. Experimental studies show RIC7 localizes to the nucleus in dark conditions and translocates to the plasma membrane upon light stimulation or ROP2 activation (Hong et al. 2016). The protein has a predicted signal peptide and transmembrane domain consistent with secretory pathway/membrane targeting rather than chloroplast import.
Reason: The computational prediction of chloroplast localization is not supported by any experimental evidence. Direct confocal microscopy shows nuclear and plasma membrane localization in guard cells, with no indication of chloroplast targeting. The predicted signal peptide is more consistent with ER/plasma membrane trafficking than chloroplast import.
GO:0005515 protein binding
IPI
PMID:11752391
A genome-wide analysis of Arabidopsis Rop-interactive CRIB m...
MODIFY
Summary: PMID:11752391 (Wu et al. 2001) identified RIC7 as one of 11 RIC proteins that interact with Rop GTPases via their CRIB motif, using yeast two-hybrid screening. The with/from column indicates AT3G51300 (ROP3/ARAC1). The protein binding annotation captures this ROP GTPase interaction but is uninformative about the specific nature of the interaction.
Reason: Protein binding (GO:0005515) is too generic per GO curation guidelines. RIC7 specifically binds activated ROP GTPases through its CRIB domain. A more informative annotation would be small GTPase binding (GO:0031267), which accurately describes the ROP GTPase interaction demonstrated by yeast two-hybrid assays and confirmed by subsequent studies.
Proposed replacements: small GTPase binding
GO:0009860 pollen tube growth
IMP
PMID:11752391
A genome-wide analysis of Arabidopsis Rop-interactive CRIB m...
KEEP AS NON CORE
Summary: Wu et al. (2001) showed that overexpression of RIC7 in tobacco pollen tubes causes growth inhibition, demonstrating a role in pollen tube growth. The IMP evidence is appropriate for this overexpression phenotype. While later studies (Hong et al. 2016) established RIC7's primary biological role in guard cell stomatal regulation, the pollen tube growth involvement is experimentally supported.
Reason: The annotation is experimentally supported by pollen tube overexpression assays, but the primary characterized biological role of RIC7 is in guard cell stomatal regulation, not pollen tube growth. The pollen tube phenotype may reflect a secondary or overexpression-specific effect. The qualifier acts_upstream_of_or_within appropriately conveys uncertainty about the directness of involvement.
GO:0016324 apical plasma membrane
IDA
PMID:11752391
A genome-wide analysis of Arabidopsis Rop-interactive CRIB m...
ACCEPT
Summary: Wu et al. (2001) showed localization of RIC proteins including RIC7 in pollen tubes. The IDA evidence indicates direct observation of RIC7 at the apical plasma membrane region. This is consistent with later findings (Hong et al. 2016) showing RIC7 plasma membrane localization in guard cells upon ROP activation. In pollen tubes, the apical region is where ROP GTPases are concentrated and active.
Reason: The apical plasma membrane localization in pollen tubes is supported by IDA evidence from the original characterization study. This is consistent with RIC7's role as a ROP effector, since ROP GTPases are enriched at the pollen tube apex.
GO:0007165 signal transduction
IC
PMID:11751054
The leucine-rich repeat as a protein recognition motif.
KEEP AS NON CORE
Summary: The signal transduction annotation is inferred by curator (IC) based on protein binding evidence (with/from GO:0005515) and supported by a review on LRR protein recognition motifs (PMID:11751054, Kobe & Kajava 2001). The inference that an LRR-containing protein that binds ROP GTPases participates in signal transduction is biologically sound. RIC7 is indeed a signaling adaptor in the ROP2-RIC7-Exo70B1 pathway.
Reason: Signal transduction is correct but very broad. RIC7 specifically participates in ROP GTPase signaling that regulates stomatal movement. The annotation is not wrong but does not capture the specific signaling pathway. The IC evidence and general review reference are appropriate for this level of annotation.

Core Functions

RIC7 functions as a signaling adaptor downstream of ROP2 GTPase, binding activated (GTP-bound) ROP proteins via its CRIB domain and directly inhibiting the exocyst subunit Exo70B1. This activity negatively regulates stomatal opening by restraining vesicle trafficking to the plasma membrane in guard cells. RIC7 also modulates the kinetics of ABA-induced stomatal closure.

Supporting Evidence:
  • PMID:11752391
    we have identified 11 Arabidopsis genes encoding novel proteins, termed RICs (for Rop-interactive CRIB motif-containing proteins), that contain a CRIB (for Cdc42/Rac-interactive binding) motif required for their specific interaction with GTP-bound Rop1
  • file:ARATH/F4JLB7/F4JLB7-deep-research-falcon.md
    RIC7 functions downstream of ROP2 and inhibits the positive stomatal-opening factor Exo70B1, thereby restraining excess opening

References

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
AtSubP analysis
The leucine-rich repeat as a protein recognition motif.
  • General review on LRR domains as protein-protein interaction scaffolds; used as basis for IC inference that LRR/CRIB-containing RIC7 participates in signal transduction.
A genome-wide analysis of Arabidopsis Rop-interactive CRIB motif-containing proteins that act as Rop GTPase targets.
  • Primary study identifying all 11 RIC proteins in Arabidopsis. RIC7 was shown to interact with GTP-bound ROP1 via its CRIB motif in yeast two-hybrid assays. Overexpression in tobacco pollen tubes caused growth inhibition. RIC7 was localized to the apical plasma membrane in pollen tubes.
    "we have identified 11 Arabidopsis genes encoding novel proteins, termed RICs (for Rop-interactive CRIB motif-containing proteins), that contain a CRIB (for Cdc42/Rac-interactive binding) motif required for their specific interaction with GTP-bound Rop1"
Prediction of glycosylphosphatidylinositol-anchored proteins in Arabidopsis. A genomic analysis.
  • Genomic analysis predicting GPI-anchored proteins in Arabidopsis, including plasma membrane receptors. At4g28560/RIC7 was identified in this analysis as a predicted membrane-associated receptor-like protein.
Identification of glycosylphosphatidylinositol-anchored proteins in Arabidopsis. A proteomic and genomic analysis.
  • Follow-up proteomic validation of predicted GPI-anchored proteins in Arabidopsis. Receptor-like proteins including those at the RIC7 locus were identified among GPI-anchored protein candidates.
file:ARATH/F4JLB7/F4JLB7-deep-research-falcon.md
Falcon deep research report for Arabidopsis RIC7/F4JLB7, covering the ROP2-RIC7-Exo70B1 stomatal signaling pathway
  • Comprehensive research synthesis based on Hong et al. 2016 (New Phytologist) and reviews on ROP signaling. Establishes RIC7 as a negative regulator of stomatal movement that links ROP2 activity to Exo70B1-mediated vesicle trafficking in guard cells.

Deep Research

Falcon

(F4JLB7-deep-research-falcon.md)
Comprehensive Research Report: RIC7 (ROP-interactive CRIB motif-containing protein 7) in Arabidopsis thaliana Falcon Edison Scientific Literature 9 citations 1 artifacts 2026-06-18T17:32:20.325510

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Comprehensive Research Report: RIC7 (ROP-interactive CRIB motif-containing protein 7) in Arabidopsis thaliana

Gene and Protein Identity

RIC7 (gene locus At4g28560, UniProt accession F4JLB7) encodes a ROP-interactive CRIB motif-containing protein 7 in Arabidopsis thaliana (hong2016therop2ric7pathway pages 1-2). This protein belongs to the RIC family, a group of eleven structurally divergent proteins in Arabidopsis that function as downstream effectors of ROP (Rho of Plants) GTPases (feiguelman2018ropgtpasesstructurefunction pages 1-5). The identity of RIC7 has been confirmed through molecular studies showing it contains the characteristic CRIB (Cdc42/Rac-interactive binding) domain that mediates interaction with activated ROP proteins (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 10-13).

Protein Family and Structural Features

The RIC Family in Arabidopsis

RIC proteins represent a plant-specific family of ROP effectors characterized by the presence of a CRIB domain (feiguelman2018ropgtpasesstructurefunction pages 10-13, smokvarska2021functionofmembrane pages 1-2). The CRIB domain is a conserved sequence motif found in effector proteins that interact with GTP-bound forms of Rho-family GTPases (smokvarska2021functionofmembrane pages 1-2). In plants, CRIB domains are found not only in RIC proteins but also in certain ROPGAPs (ROP GTPase-activating proteins), where they enhance binding between ROPs and ROPGAPs and are required for subcellular localization (feiguelman2018ropgtpasesstructurefunction pages 10-13).

Arabidopsis possesses eleven RIC proteins (RIC1-11) that have evolved to control various ROP GTPase-dependent pathways through functional diversification (hong2016therop2ric7pathway pages 1-2). While some RIC family members like RIC1, RIC3, and RIC4 regulate cytoskeletal organization during processes such as pollen tube growth and cell morphogenesis (feiguelman2018ropgtpasesstructurefunction pages 1-5), RIC7 represents a specialized branch involved in guard cell signaling rather than the cytoskeletal functions typical of other RIC proteins (feiguelman2018ropgtpasesstructurefunction pages 1-5).

Domain Architecture

The defining feature of RIC7 is its CRIB motif, which enables direct interaction with activated (GTP-bound) ROP proteins (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 10-13). This domain is essential for RIC7 to function as a molecular effector that transduces ROP signaling to downstream cellular processes. The CRIB domain binds preferentially to the active conformation of ROPs, ensuring signal specificity (smokvarska2021functionofmembrane pages 1-2).

Primary Molecular Function

Role as a Negative Regulator of Stomatal Movement

The primary function of RIC7 is to negatively regulate stomatal movements in Arabidopsis guard cells (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 3-4). Genetic and biochemical studies have established RIC7 as a key component that fine-tunes both light-induced stomatal opening and abscisic acid (ABA)-induced stomatal closure.

Knockout studies using the ric7-1 mutant (which harbors a T-DNA insertion in the second intron) demonstrated that loss of RIC7 function leads to accelerated light-induced stomatal opening (hong2016therop2ric7pathway pages 3-4). After 1-2 hours of light irradiation, ric7-1 mutant stomata opened approximately 18% more widely than wild-type stomata (hong2016therop2ric7pathway pages 3-4). Conversely, plants overexpressing RIC7 under the control of the 35S promoter exhibited slower light-induced stomatal opening compared to wild-type plants (hong2016therop2ric7pathway pages 3-4). Complementation lines expressing genomic RIC7 driven by its native promoter in the ric7-1 background restored wild-type stomatal opening kinetics, confirming that the rapid opening phenotype was due to RIC7 loss of function (hong2016therop2ric7pathway pages 3-4).

RIC7 also suppresses ABA-induced stomatal closure. The ric7-1 knockout mutant exhibited faster ABA-induced stomatal closure than wild-type, with differences evident as early as 30 minutes after ABA treatment (hong2016therop2ric7pathway pages 3-4). RIC7 overexpression lines showed slower ABA-induced closure (hong2016therop2ric7pathway pages 3-4). These results indicate that RIC7 functions as a negative regulator of both stomatal opening and closure, similar to its upstream regulator ROP2 (hong2016therop2ric7pathway pages 8-9).

Mechanistic Basis: The ROP2-RIC7-Exo70B1 Signaling Module

RIC7 does not function as a substrate-specific enzyme or transporter. Instead, it operates as a signaling adaptor protein that links ROP2 GTPase activity to the vesicle trafficking machinery required for stomatal movement (hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10).

Interaction with ROP2: RIC7 functions downstream of ROP2, a plant Rho-type GTPase that negatively regulates stomatal movements (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 8-9). Previous work established that RIC7 binds to activated ROP2 in vitro, positioning it as a ROP2 effector (hong2016therop2ric7pathway pages 1-2). The interaction between RIC7 and active ROP2 is mediated by the CRIB domain (hong2016therop2ric7pathway pages 1-2).

Direct Binding to Exo70B1: A critical discovery was the identification of Exo70B1 (exocyst subunit Exo70 family protein B1) as a direct interaction partner of RIC7 (hong2016therop2ric7pathway pages 4-5). Multiple experimental approaches confirmed this interaction:

  • Yeast two-hybrid assays demonstrated that RIC7 binds to Exo70B1 (hong2016therop2ric7pathway pages 4-5)
  • In vitro pull-down assays showed that GST-fused Exo70B1 directly pulls down MBP-fused RIC7, whereas free GST does not (hong2016therop2ric7pathway pages 4-5)
  • Bimolecular fluorescence complementation (BiFC) assays in tobacco leaves confirmed the RIC7-Exo70B1 interaction in vivo (hong2016therop2ric7pathway pages 4-5)

Importantly, BiFC assays revealed that Exo70B1 does not bind directly to ROP2 (hong2016therop2ric7pathway pages 4-5), indicating that RIC7 serves as a linker protein connecting ROP2 to the exocyst machinery. This differs from yeast and animal systems where Rho GTPases often bind directly to Exo70 proteins (hong2016therop2ric7pathway pages 8-9). The requirement for a linker protein may reflect the absence of conserved C-terminal Rho-binding motifs in plant Exo70 proteins (hong2016therop2ric7pathway pages 8-9).

Functional Relationship with Exo70B1: RIC7 and Exo70B1 have opposing functions in stomatal regulation. While RIC7 is a negative regulator of light-induced stomatal opening, Exo70B1 is a positive regulator (hong2016therop2ric7pathway pages 5-7, hong2016therop2ric7pathway pages 9-10). The exo70b1-1 and exo70b1-2 knockout mutants showed retarded light-induced stomatal opening, with stomata opening approximately 14% less than wild-type after 1 hour of light (hong2016therop2ric7pathway pages 5-7). Genetic epistasis analysis using the ric7-1/exo70b1-1 double knockout mutant demonstrated that the double mutant phenotype resembled exo70b1-1 rather than ric7-1, indicating that Exo70B1 functions downstream of RIC7 (hong2016therop2ric7pathway pages 7-8).

The proposed mechanism is that RIC7 binding inhibits Exo70B1 function, thereby restraining vesicle trafficking and limiting the rate of stomatal opening (hong2016therop2ric7pathway pages 9-10). This creates a brake mechanism that prevents excessive stomatal opening and associated water loss while still allowing efficient COโ‚‚ uptake for photosynthesis (hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10).

Subcellular Localization and Dynamic Regulation

RIC7 exhibits stimulus-dependent subcellular localization, which is critical for its regulatory function (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 5-7).

Localization Under Dark Conditions: In guard cells maintained in darkness, CFP-tagged RIC7 localizes predominantly to the nucleus (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 5-7). Under these conditions, YFP-tagged Exo70B1 shows a punctate pattern in the cytosol and nucleus (hong2016therop2ric7pathway pages 7-8).

Light-Induced Relocalization: Upon light irradiation (2 hours of white light), a subset of RIC7 relocates from the nucleus to the plasma membrane region, where it co-localizes with Exo70B1 (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 5-7). Fluorescence intensity profiles confirmed co-localization of YFP-Exo70B1 and CFP-RIC7 at the plasma membrane in light-treated guard cells (hong2016therop2ric7pathway pages 7-8).

ROP2 Activity-Dependent Localization: The relocalization of RIC7 to the plasma membrane is dependent on ROP2 activity. When co-expressed with constitutively active ROP2 (CA-ROP2), RIC7 and Exo70B1 both localize to the plasma membrane region even in the absence of light (hong2016therop2ric7pathway pages 7-8). In contrast, when co-expressed with a dominant-negative ROP2 (DN-ROP2), RIC7 remains primarily nuclear and Exo70B1 maintains its punctate cytoplasmic/nuclear distribution without plasma membrane enrichment (hong2016therop2ric7pathway pages 7-8). These findings indicate that light-mediated ROP2 activation triggers the recruitment of both RIC7 and Exo70B1 to the plasma membrane, where RIC7 can then modulate Exo70B1 function (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 9-10).

Biological Processes and Signaling Pathways

Guard Cell Function and Gas Exchange

RIC7 is highly expressed in stomatal guard cells, as demonstrated by RIC7 promoter::ฮฒ-glucuronidase reporter analysis (hong2016therop2ric7pathway pages 3-4). Guard cells regulate stomatal apertures to balance COโ‚‚ uptake for photosynthesis against water loss through transpiration. Stomatal opening requires increases in guard cell volume driven by ion accumulation and water influx, necessitating expansion of the plasma membrane and reorganization of the vacuolar membrane (hong2016therop2ric7pathway pages 9-10).

The ROP2 Signaling Pathway in Guard Cells

ROP2 (also known as RAC2) is a member of the plant-specific ROP subfamily of Rho GTPases and functions as a negative regulator of stomatal movements (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 1-5). Like all small GTPases, ROP proteins cycle between an inactive GDP-bound state and an active GTP-bound state. This cycle is regulated by guanine nucleotide exchange factors (GEFs) that promote GDP release and GTP binding, and GTPase-activating proteins (GAPs) that enhance GTP hydrolysis (feiguelman2018ropgtpasesstructurefunction pages 1-5, smokvarska2021functionofmembrane pages 1-2).

In the context of stomatal regulation, light perception by phototropins and chlorophylls activates signaling pathways that ultimately modulate ROP2 activity (hong2016therop2ric7pathway pages 9-10). The current model proposes that light-induced stomatal opening activates positive regulatory pathways but simultaneously engages negative regulatory mechanisms, including the ROP2-RIC7-Exo70B1 pathway, to prevent excessive opening (hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10).

Vesicle Trafficking and Membrane Remodeling

The functional interaction between RIC7 and Exo70B1 implicates vesicle trafficking as a critical regulatory target in stomatal movement (hong2016therop2ric7pathway pages 9-10). Exo70B1 is a component of the exocyst complex, an eight-subunit complex (Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84) that mediates tethering and spatial targeting of post-Golgi vesicles to the plasma membrane prior to fusion (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 2-3).

In guard cells, Exo70B1 co-localizes with Exo84B, another exocyst component, supporting its role in exocyst function (hong2016therop2ric7pathway pages 7-8). However, Exo70B1 shows minimal co-localization with Golgi or trans-Golgi network/early endosome (TGN/EE) markers (only 3-4% overlap with Golgi marker ST-GFP and 11-12% with TGN/EE marker YFP-SYP61), suggesting involvement in non-conventional trafficking pathways or exocyst-positive organelle (EXPO)-mediated exocytosis (hong2016therop2ric7pathway pages 8-9).

The proposed mechanism is that Exo70B1 facilitates the trafficking of ion transporters and membrane materials to the plasma membrane and vacuolar membrane during stomatal opening (hong2016therop2ric7pathway pages 9-10). RIC7 binding to Exo70B1 inhibits this trafficking function, thereby limiting the rate of membrane expansion and ion transporter delivery, which in turn restrains the rate of stomatal opening (hong2016therop2ric7pathway pages 9-10). This provides a mechanism for plants to fine-tune stomatal aperture in response to environmental conditions.

Integration with ABA Signaling

RIC7 also modulates ABA-induced stomatal closure, although the precise mechanism differs from its role in light-induced opening (hong2016therop2ric7pathway pages 3-4, hong2016therop2ric7pathway pages 8-9). The observation that ric7 mutants exhibit faster ABA-induced closure suggests that RIC7 normally acts to dampen or slow the closure response (hong2016therop2ric7pathway pages 3-4). This may reflect a general role for RIC7 in modulating the kinetics of membrane trafficking events during stomatal movements, regardless of the directional stimulus.

Previous work has established that ROP proteins and ABA signaling form negative feedback loops (feiguelman2018ropgtpasesstructurefunction pages 1-5). ABA signaling suppresses ROP activation by promoting degradation of ROPGEFs, while ROP signaling suppresses ABA responses by physical interactions between ROPs (particularly ROP10 and ROP11) and ABI1/ABI2 PP2C phosphatases, negative regulators of ABA signaling (feiguelman2018ropgtpasesstructurefunction pages 1-5). RIC7's role in this network likely involves transducing ROP2 signals that modulate the kinetics of stomatal responses to both positive (light) and negative (ABA) stimuli.

Experimental Evidence

The characterization of RIC7 is supported by multiple lines of experimental evidence employing diverse methodologies:

Genetic Evidence:
- T-DNA insertion mutant ric7-1 (GK_062G02) with insertion in the second intron (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 3-4)
- RIC7 overexpression lines (OX1, OX3, OX8) under 35S promoter control (hong2016therop2ric7pathway pages 3-4)
- Complementation lines expressing genomic RIC7 under native promoter in ric7-1 background (hong2016therop2ric7pathway pages 3-4)
- Double mutant ric7-1/exo70b1-1 for epistasis analysis (hong2016therop2ric7pathway pages 7-8)

Biochemical Evidence:
- Yeast two-hybrid screens identifying Exo70B1 as RIC7-interacting protein (hong2016therop2ric7pathway pages 4-5)
- In vitro GST pull-down assays demonstrating direct RIC7-Exo70B1 interaction (hong2016therop2ric7pathway pages 4-5)
- Quantitative RT-PCR confirming gene expression levels in mutants and transgenics (hong2016therop2ric7pathway pages 2-3)

Cell Biological Evidence:
- Bimolecular fluorescence complementation (BiFC) assays in Nicotiana benthamiana confirming RIC7-Exo70B1 interaction in vivo (hong2016therop2ric7pathway pages 4-5)
- Transient expression of fluorescently tagged proteins in Vicia faba guard cells via particle bombardment (hong2016therop2ric7pathway pages 2-3, hong2016therop2ric7pathway pages 7-8)
- Confocal microscopy analysis of subcellular localization under different conditions (dark, light, with CA-ROP2, with DN-ROP2) (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 5-7)
- Co-localization studies with organelle markers (hong2016therop2ric7pathway pages 8-9)

Physiological Evidence:
- Stomatal aperture measurements showing altered light-induced opening kinetics in ric7 mutants and overexpression lines (hong2016therop2ric7pathway pages 3-4)
- ABA-induced stomatal closure assays demonstrating RIC7's role in closure kinetics (hong2016therop2ric7pathway pages 3-4)
- Promoter::ฮฒ-glucuronidase reporter analysis revealing RIC7 expression patterns (hong2016therop2ric7pathway pages 3-4)

Comparison with Other RIC Family Members

The RIC family exhibits functional diversification, with different members regulating distinct cellular processes (feiguelman2018ropgtpasesstructurefunction pages 1-5). While RIC7 is specialized for guard cell signaling and vesicle trafficking regulation, other RIC proteins have well-characterized roles in cytoskeletal regulation:

  • RIC1 regulates microtubule-mediated cell morphogenesis and localizes to the pollen tube tip (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 1-5)
  • RIC3 and RIC4 regulate cortical actin filament organization in growing pollen tubes (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 1-5)
  • RIC171 in barley (a RIC family ortholog) is involved in pathogen defense responses, where it accumulates at sites of fungal attack (feiguelman2018ropgtpasesstructurefunction pages 20-23)

This functional diversity reflects the evolution of the RIC family to mediate different downstream outputs of ROP signaling in different cellular contexts, despite sharing the common CRIB domain that enables ROP interaction (feiguelman2018ropgtpasesstructurefunction pages 1-5).

Current Understanding and Knowledge Gaps

The characterization of RIC7 published by Hong et al. in 2016 remains the most comprehensive functional study of this protein (hong2016therop2ric7pathway pages 1-2). Recent reviews (2020-2022) on ROP signaling pathways provide broader context for understanding RIC proteins as ROP effectors (feiguelman2018ropgtpasesstructurefunction pages 1-5, smokvarska2021functionofmembrane pages 1-2), but RIC7-specific studies remain limited compared to other RIC family members like RIC1, RIC3, and RIC4.

Key remaining questions include:
1. The precise molecular mechanism by which RIC7 binding inhibits Exo70B1 function
2. Whether RIC7 interacts with other exocyst components or trafficking machinery
3. The structural basis of RIC7-Exo70B1 interaction
4. Potential roles of RIC7 in other cell types beyond guard cells
5. The evolutionary origin and conservation of RIC7 function in other plant species

Summary

Category Findings for RIC7 (Arabidopsis thaliana) Evidence / citation
Gene / protein identity RIC7; Arabidopsis locus At4g28560; UniProt accession F4JLB7; protein name ROP-interactive CRIB motif-containing protein 7; organism Arabidopsis thaliana. Identity is consistent with the Arabidopsis RIC family described as CRIB-domain ROP effectors. (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 1-5)
Family / defining features RIC7 belongs to the RIC (ROP-interactive CRIB motif-containing) family, a set of structurally divergent Arabidopsis ROP effectors that bind activated ROP GTPases via a CRIB domain. Arabidopsis contains 11 RIC proteins. (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 10-13, smokvarska2021functionofmembrane pages 1-2)
Current domain understanding The defining experimentally supported feature is the CRIB motif/domain, which mediates interaction with active ROPs in plant RIC proteins. More broadly, CRIB-containing proteins function as downstream effectors of Rho/ROP-family GTPases. (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 10-13, smokvarska2021functionofmembrane pages 1-2)
Primary molecular function RIC7 is a negative regulator of stomatal movement, especially light-induced stomatal opening; it also suppresses ABA-induced stomatal closure. Mechanistically, it acts downstream of ROP2 and inhibits the positive stomatal-opening factor Exo70B1, thereby restraining excess opening. (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 3-4, hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10)
Pathway position In the best-supported pathway, active ROP2 โ†’ RIC7 โ†’ Exo70B1 inhibition, with RIC7 functioning as a linker/effector that transduces ROP2 signaling to vesicle-trafficking machinery in guard cells. (hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10)
Direct interaction partners ROP2: prior work placed RIC7 downstream of active ROP2; Exo70B1: directly binds RIC7 in yeast two-hybrid, in vitro pull-down, and BiFC assays. Exo70B1 did not bind ROP2 directly in the reported BiFC assay, supporting a linker role for RIC7. (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 4-5, hong2016therop2ric7pathway pages 8-9)
Positive / opposing functional partner Exo70B1 is a positive regulator of light-induced stomatal opening, whereas RIC7 is negative; the exo70b1 phenotype is epistatic to ric7 in double mutants, supporting Exo70B1 as a downstream target of RIC7. (hong2016therop2ric7pathway pages 5-7, hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 9-10)
Subcellular localization In guard cells under dark conditions, RIC7 is localized mostly to the nucleus; under light or with constitutively active ROP2, a subset of RIC7 relocates to the plasma membrane region, where it co-localizes with Exo70B1. This dynamic relocalization is consistent with stimulus-dependent signaling. (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 5-7)
Localization context of partner Exo70B1 localizes to the plasma membrane region, nucleus, and cytosolic puncta; it co-localizes with Exo84B but shows little co-localization with Golgi or TGN/EE markers, suggesting association with exocyst-related / noncanonical trafficking compartments relevant to RIC7 action. (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10)
Tissue / expression context RIC7 promoter activity is detected broadly in young tissues and is high in stomatal guard cells, supporting a direct role in guard-cell physiology. (hong2016therop2ric7pathway pages 3-4)
Biological processes implicated Guard-cell signaling, stomatal opening, ABA-responsive stomatal closure, and likely vesicle trafficking / membrane remodeling needed for stomatal movement. The proposed mechanism is inhibition of exocyst-mediated trafficking required for rapid opening. (hong2016therop2ric7pathway pages 1-2, hong2016therop2ric7pathway pages 9-10)
Functional model Light activates stomatal opening pathways but also a brake mechanism: ROP2 recruits RIC7 and Exo70B1 to the plasma membrane region; RIC7 binds Exo70B1 and inhibits its function, fine-tuning opening kinetics and preventing excessive water loss. (hong2016therop2ric7pathway pages 7-8, hong2016therop2ric7pathway pages 8-9, hong2016therop2ric7pathway pages 9-10)
Genetic evidence ric7-1 knockout shows faster light-induced opening and faster ABA-induced closure than wild type; RIC7 complementation restores wild-type behavior; RIC7 overexpression slows both responses. (hong2016therop2ric7pathway pages 3-4)
Biochemical / cell-biological evidence Evidence includes yeast two-hybrid, in vitro GST pull-down, BiFC, transient fluorescent localization in Vicia faba guard cells, and Arabidopsis mutant / complementation / overexpression analysis. Together these support direct RIC7โ€“Exo70B1 interaction and dynamic localization. (hong2016therop2ric7pathway pages 4-5, hong2016therop2ric7pathway pages 5-7, hong2016therop2ric7pathway pages 7-8)
Broader family context RIC proteins are diverse ROP effectors. Other Arabidopsis RICs regulate actin and microtubule organization and polarized growth, indicating that RIC7 likely represents a specialized branch of ROP effector biology tied to guard-cell membrane trafficking rather than the canonical pollen-tube cytoskeletal outputs of RIC1/3/4. (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 1-5, smokvarska2021functionofmembrane pages 1-2)
Key references summarized in evidence base Primary experimental study: Hong et al., 2016, New Phytologist on the ROP2โ€“RIC7โ€“Exo70B1 pathway in stomatal opening. Conceptual background from ROP signaling reviews in Plant Physiology 2018 and Plant Physiology 2021. (hong2016therop2ric7pathway pages 1-2, feiguelman2018ropgtpasesstructurefunction pages 1-5, smokvarska2021functionofmembrane pages 1-2)

Table: This table compiles the verified identity, molecular function, localization, pathway role, and supporting evidence for Arabidopsis RIC7. It is useful as a concise annotation-ready summary centered on the experimentally supported ROP2-RIC7-Exo70B1 guard-cell signaling module.

RIC7 (At4g28560, UniProt F4JLB7) is a ROP-interactive CRIB motif-containing protein that functions as a negative regulator of stomatal movements in Arabidopsis thaliana. As a member of the eleven-protein RIC family, RIC7 serves as a downstream effector of ROP2 GTPase, transducing signals to the vesicle trafficking machinery through direct interaction with exocyst subunit Exo70B1. RIC7 exhibits dynamic subcellular localization, moving from the nucleus to the plasma membrane region in response to light stimulation or ROP2 activation. By binding to and inhibiting Exo70B1, RIC7 restrains excessive stomatal opening, providing a brake mechanism that balances COโ‚‚ uptake against water loss. This function has been validated through comprehensive genetic, biochemical, and cell biological approaches, including analysis of knockout mutants, overexpression lines, protein-protein interaction assays, and subcellular localization studies. RIC7 represents a specialized branch of RIC protein evolution, diverging from the cytoskeletal regulatory functions of other family members to perform a unique role in guard cell physiology and gas exchange regulation.

Primary Reference: Hong, D., Jeon, B.W., Kim, S.Y., Hwang, J.-U., & Lee, Y. (2016). The ROP2-RIC7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit Exo70B1 in Arabidopsis. New Phytologist, 209(2), 624-635. doi:10.1111/nph.13625

Publication Date: October 2015 (published online), January 2016 (print)

DOI/URL: https://doi.org/10.1111/nph.13625

References

  1. (hong2016therop2ric7pathway pages 1-2): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  2. (feiguelman2018ropgtpasesstructurefunction pages 1-5): Gil Feiguelman, Ying Fu, and Shaul Yalovsky. Rop gtpases structure-function and signaling pathways1[open]. Plant Physiology, 176:57-79, Nov 2018. URL: https://doi.org/10.1104/pp.17.01415, doi:10.1104/pp.17.01415. This article has 227 citations and is from a highest quality peer-reviewed journal.

  3. (feiguelman2018ropgtpasesstructurefunction pages 10-13): Gil Feiguelman, Ying Fu, and Shaul Yalovsky. Rop gtpases structure-function and signaling pathways1[open]. Plant Physiology, 176:57-79, Nov 2018. URL: https://doi.org/10.1104/pp.17.01415, doi:10.1104/pp.17.01415. This article has 227 citations and is from a highest quality peer-reviewed journal.

  4. (smokvarska2021functionofmembrane pages 1-2): Marija Smokvarska, Yvon Jaillais, and Alexandre Martiniรจre. Function of membrane domains in rho-of-plant signaling. Plant Physiology, 185:663-681, Jan 2021. URL: https://doi.org/10.1093/plphys/kiaa082, doi:10.1093/plphys/kiaa082. This article has 54 citations and is from a highest quality peer-reviewed journal.

  5. (hong2016therop2ric7pathway pages 3-4): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  6. (hong2016therop2ric7pathway pages 8-9): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  7. (hong2016therop2ric7pathway pages 9-10): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  8. (hong2016therop2ric7pathway pages 4-5): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  9. (hong2016therop2ric7pathway pages 5-7): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  10. (hong2016therop2ric7pathway pages 7-8): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  11. (hong2016therop2ric7pathway pages 2-3): Daewoong Hong, Byeong Wook Jeon, Soo Young Kim, Jaeโ€Ung Hwang, and Youngsook Lee. The rop2-ric7 pathway negatively regulates light-induced stomatal opening by inhibiting exocyst subunit exo70b1 in arabidopsis. The New phytologist, 209 2:624-35, Oct 2016. URL: https://doi.org/10.1111/nph.13625, doi:10.1111/nph.13625. This article has 72 citations.

  12. (feiguelman2018ropgtpasesstructurefunction pages 20-23): Gil Feiguelman, Ying Fu, and Shaul Yalovsky. Rop gtpases structure-function and signaling pathways1[open]. Plant Physiology, 176:57-79, Nov 2018. URL: https://doi.org/10.1104/pp.17.01415, doi:10.1104/pp.17.01415. This article has 227 citations and is from a highest quality peer-reviewed journal.

Artifacts

Citations

  1. feiguelman2018ropgtpasesstructurefunction pages 1-5
  2. smokvarska2021functionofmembrane pages 1-2
  3. feiguelman2018ropgtpasesstructurefunction pages 10-13
  4. feiguelman2018ropgtpasesstructurefunction pages 20-23
  5. open
  6. https://doi.org/10.1111/nph.13625
  7. https://doi.org/10.1111/nph.13625,
  8. https://doi.org/10.1104/pp.17.01415,
  9. https://doi.org/10.1093/plphys/kiaa082,

๐Ÿ“„ View Raw YAML

id: F4JLB7
gene_symbol: RIC7
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:3702
  label: Arabidopsis thaliana
description: >-
  RIC7 (ROP-interactive CRIB motif-containing protein 7; locus At4g28560; UniProt
  F4JLB7) is a member of the 11-protein RIC family in Arabidopsis thaliana. RIC
  proteins are plant-specific effectors of ROP (Rho of Plants) GTPases, defined by
  a conserved CRIB (Cdc42/Rac-interactive binding) domain that mediates
  preferential binding to the GTP-bound (active) form of ROP proteins. RIC7 also
  contains leucine-rich repeat (LRR) domains, placing it in the receptor-like
  protein (RLP) family. RIC7 functions as a signaling adaptor downstream of ROP2,
  negatively regulating both light-induced stomatal opening and ABA-induced
  stomatal closure in guard cells. Upon light-stimulated ROP2 activation, RIC7
  translocates from the nucleus to the plasma membrane, where it directly binds
  and inhibits the exocyst subunit Exo70B1, thereby restraining vesicle
  trafficking required for rapid stomatal aperture changes. The ROP2-RIC7-Exo70B1
  signaling module provides a brake mechanism that fine-tunes stomatal kinetics to
  balance CO2 uptake against water loss. RIC7 is highly expressed in guard cells
  and has also been implicated in pollen tube growth through its interaction with
  ROP GTPases.
existing_annotations:
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: is_active_in
  review:
    summary: >-
      RIC7 dynamically localizes to the plasma membrane region in guard cells upon
      light stimulation or co-expression with constitutively active ROP2 (Hong et
      al. 2016). The IBA annotation from phylogenetic inference is consistent with
      the experimentally demonstrated plasma membrane localization in guard cells.
    action: ACCEPT
    reason: >-
      Plasma membrane localization of RIC7 is well supported by confocal microscopy
      of fluorescently tagged RIC7 in Vicia faba guard cells, showing
      light-dependent and ROP2-activity-dependent relocalization to the plasma
      membrane. The phylogenetic inference is consistent with experimental data.
- term:
    id: GO:0038023
    label: signaling receptor activity
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: >-
      RIC7 is not a signaling receptor per se. It is an intracellular signaling
      adaptor/effector that binds activated ROP GTPases via its CRIB domain and
      transduces their signals to downstream targets. While it contains LRR domains
      typical of receptor-like proteins, there is no evidence that RIC7 itself
      perceives extracellular ligands or has signaling receptor activity. The IBA
      annotation likely propagated from receptor-like protein family members that do
      have receptor activity, but RIC7 lacks a kinase domain and functions as a
      cytoplasmic effector rather than a receptor.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      The phylogenetic annotation appears to have propagated receptor activity from
      LRR receptor-like kinase or receptor-like protein family members. However,
      RIC7 is functionally characterized as an intracellular ROP GTPase effector,
      not a signaling receptor. Its CRIB motif mediates intracellular
      protein-protein interactions with activated ROP2, not extracellular ligand
      perception. Small GTPase binding (GO:0031267) would be more appropriate for
      this protein.
- term:
    id: GO:0016020
    label: membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: >-
      UniProt computationally predicts F4JLB7 as a single-pass type I membrane
      protein based on ARBA analysis. The protein does contain a predicted signal
      peptide and transmembrane domain. Experimental evidence supports plasma
      membrane localization in guard cells, so the general membrane annotation is
      consistent but less informative than the more specific plasma membrane
      annotation.
    action: KEEP_AS_NON_CORE
    reason: >-
      The annotation is correct but overly broad. The protein localizes to the
      plasma membrane specifically, and more specific annotations exist
      (GO:0005886, GO:0016324). This IEA annotation is subsumed by those.
- term:
    id: GO:0016020
    label: membrane
  evidence_type: TAS
  original_reference_id: PMID:12068095
  qualifier: located_in
  review:
    summary: >-
      PMID:12068095 (Borner et al. 2002) is a genomic analysis predicting
      GPI-anchored proteins in Arabidopsis. The paper identified RIC7 (At4g28560)
      as a predicted plasma membrane receptor-like protein. The membrane
      localization is consistent with this computational prediction and with
      subsequent experimental evidence.
    action: KEEP_AS_NON_CORE
    reason: >-
      The annotation is broadly correct but nonspecific. More informative
      localization terms (plasma membrane, apical plasma membrane) are available
      from other annotations. The TAS evidence is based on computational
      predictions of GPI-anchoring and membrane association rather than direct
      experimental demonstration.
- term:
    id: GO:0016020
    label: membrane
  evidence_type: TAS
  original_reference_id: PMID:12805588
  qualifier: located_in
  review:
    summary: >-
      PMID:12805588 (Borner et al. 2003) is a proteomic and genomic analysis of
      GPI-anchored proteins in Arabidopsis that identified receptor-like proteins
      including those at the At4g28560 locus. The membrane annotation is consistent
      with these findings.
    action: KEEP_AS_NON_CORE
    reason: >-
      Same rationale as the other membrane annotations. The annotation is correct
      but subsumed by more specific plasma membrane and apical plasma membrane
      annotations from direct experimental evidence.
- term:
    id: GO:0009507
    label: chloroplast
  evidence_type: ISM
  original_reference_id: GO_REF:0000122
  qualifier: located_in
  review:
    summary: >-
      The chloroplast localization is based on AtSubP computational prediction
      (ISM evidence). There is no experimental evidence supporting chloroplast
      localization for RIC7. Experimental studies show RIC7 localizes to the
      nucleus in dark conditions and translocates to the plasma membrane upon light
      stimulation or ROP2 activation (Hong et al. 2016). The protein has a
      predicted signal peptide and transmembrane domain consistent with secretory
      pathway/membrane targeting rather than chloroplast import.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      The computational prediction of chloroplast localization is not supported by
      any experimental evidence. Direct confocal microscopy shows nuclear and
      plasma membrane localization in guard cells, with no indication of
      chloroplast targeting. The predicted signal peptide is more consistent with
      ER/plasma membrane trafficking than chloroplast import.
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:11752391
  qualifier: enables
  review:
    summary: >-
      PMID:11752391 (Wu et al. 2001) identified RIC7 as one of 11 RIC proteins
      that interact with Rop GTPases via their CRIB motif, using yeast two-hybrid
      screening. The with/from column indicates AT3G51300 (ROP3/ARAC1). The
      protein binding annotation captures this ROP GTPase interaction but is
      uninformative about the specific nature of the interaction.
    action: MODIFY
    reason: >-
      Protein binding (GO:0005515) is too generic per GO curation guidelines. RIC7
      specifically binds activated ROP GTPases through its CRIB domain. A more
      informative annotation would be small GTPase binding (GO:0031267), which
      accurately describes the ROP GTPase interaction demonstrated by yeast
      two-hybrid assays and confirmed by subsequent studies.
    proposed_replacement_terms:
    - id: GO:0031267
      label: small GTPase binding
- term:
    id: GO:0009860
    label: pollen tube growth
  evidence_type: IMP
  original_reference_id: PMID:11752391
  qualifier: acts_upstream_of_or_within
  review:
    summary: >-
      Wu et al. (2001) showed that overexpression of RIC7 in tobacco pollen tubes
      causes growth inhibition, demonstrating a role in pollen tube growth. The IMP
      evidence is appropriate for this overexpression phenotype. While later studies
      (Hong et al. 2016) established RIC7's primary biological role in guard cell
      stomatal regulation, the pollen tube growth involvement is experimentally
      supported.
    action: KEEP_AS_NON_CORE
    reason: >-
      The annotation is experimentally supported by pollen tube overexpression
      assays, but the primary characterized biological role of RIC7 is in guard
      cell stomatal regulation, not pollen tube growth. The pollen tube phenotype
      may reflect a secondary or overexpression-specific effect. The qualifier
      acts_upstream_of_or_within appropriately conveys uncertainty about the
      directness of involvement.
- term:
    id: GO:0016324
    label: apical plasma membrane
  evidence_type: IDA
  original_reference_id: PMID:11752391
  qualifier: located_in
  review:
    summary: >-
      Wu et al. (2001) showed localization of RIC proteins including RIC7 in
      pollen tubes. The IDA evidence indicates direct observation of RIC7 at the
      apical plasma membrane region. This is consistent with later findings (Hong
      et al. 2016) showing RIC7 plasma membrane localization in guard cells upon
      ROP activation. In pollen tubes, the apical region is where ROP GTPases are
      concentrated and active.
    action: ACCEPT
    reason: >-
      The apical plasma membrane localization in pollen tubes is supported by IDA
      evidence from the original characterization study. This is consistent with
      RIC7's role as a ROP effector, since ROP GTPases are enriched at the pollen
      tube apex.
- term:
    id: GO:0007165
    label: signal transduction
  evidence_type: IC
  original_reference_id: PMID:11751054
  qualifier: acts_upstream_of_or_within
  review:
    summary: >-
      The signal transduction annotation is inferred by curator (IC) based on
      protein binding evidence (with/from GO:0005515) and supported by a review on
      LRR protein recognition motifs (PMID:11751054, Kobe & Kajava 2001). The
      inference that an LRR-containing protein that binds ROP GTPases participates
      in signal transduction is biologically sound. RIC7 is indeed a signaling
      adaptor in the ROP2-RIC7-Exo70B1 pathway.
    action: KEEP_AS_NON_CORE
    reason: >-
      Signal transduction is correct but very broad. RIC7 specifically participates
      in ROP GTPase signaling that regulates stomatal movement. The annotation is
      not wrong but does not capture the specific signaling pathway. The IC
      evidence and general review reference are appropriate for this level of
      annotation.
references:
- 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:0000122
  title: AtSubP analysis
  findings: []
- id: PMID:11751054
  title: The leucine-rich repeat as a protein recognition motif.
  findings:
  - statement: >-
      General review on LRR domains as protein-protein interaction scaffolds; used
      as basis for IC inference that LRR/CRIB-containing RIC7 participates in
      signal transduction.
- id: PMID:11752391
  title: >-
    A genome-wide analysis of Arabidopsis Rop-interactive CRIB motif-containing
    proteins that act as Rop GTPase targets.
  findings:
  - statement: >-
      Primary study identifying all 11 RIC proteins in Arabidopsis. RIC7 was shown
      to interact with GTP-bound ROP1 via its CRIB motif in yeast two-hybrid
      assays. Overexpression in tobacco pollen tubes caused growth inhibition.
      RIC7 was localized to the apical plasma membrane in pollen tubes.
    supporting_text: >-
      we have identified 11 Arabidopsis genes encoding novel proteins, termed RICs
      (for Rop-interactive CRIB motif-containing proteins), that contain a CRIB
      (for Cdc42/Rac-interactive binding) motif required for their specific
      interaction with GTP-bound Rop1
- id: PMID:12068095
  title: >-
    Prediction of glycosylphosphatidylinositol-anchored proteins in Arabidopsis.
    A genomic analysis.
  findings:
  - statement: >-
      Genomic analysis predicting GPI-anchored proteins in Arabidopsis, including
      plasma membrane receptors. At4g28560/RIC7 was identified in this analysis as
      a predicted membrane-associated receptor-like protein.
- id: PMID:12805588
  title: >-
    Identification of glycosylphosphatidylinositol-anchored proteins in
    Arabidopsis. A proteomic and genomic analysis.
  findings:
  - statement: >-
      Follow-up proteomic validation of predicted GPI-anchored proteins in
      Arabidopsis. Receptor-like proteins including those at the RIC7 locus were
      identified among GPI-anchored protein candidates.
- id: file:ARATH/F4JLB7/F4JLB7-deep-research-falcon.md
  title: >-
    Falcon deep research report for Arabidopsis RIC7/F4JLB7, covering the
    ROP2-RIC7-Exo70B1 stomatal signaling pathway
  findings:
  - statement: >-
      Comprehensive research synthesis based on Hong et al. 2016 (New Phytologist)
      and reviews on ROP signaling. Establishes RIC7 as a negative regulator of
      stomatal movement that links ROP2 activity to Exo70B1-mediated vesicle
      trafficking in guard cells.
core_functions:
- description: >-
    RIC7 functions as a signaling adaptor downstream of ROP2 GTPase, binding
    activated (GTP-bound) ROP proteins via its CRIB domain and directly inhibiting
    the exocyst subunit Exo70B1. This activity negatively regulates stomatal
    opening by restraining vesicle trafficking to the plasma membrane in guard
    cells. RIC7 also modulates the kinetics of ABA-induced stomatal closure.
  supported_by:
  - reference_id: PMID:11752391
    supporting_text: >-
      we have identified 11 Arabidopsis genes encoding novel proteins, termed RICs
      (for Rop-interactive CRIB motif-containing proteins), that contain a CRIB
      (for Cdc42/Rac-interactive binding) motif required for their specific
      interaction with GTP-bound Rop1
  - reference_id: file:ARATH/F4JLB7/F4JLB7-deep-research-falcon.md
    supporting_text: >-
      RIC7 functions downstream of ROP2 and inhibits the positive stomatal-opening
      factor Exo70B1, thereby restraining excess opening
  molecular_function:
    id: GO:0031267
    label: small GTPase binding
  directly_involved_in:
  - id: GO:1902457
    label: negative regulation of stomatal opening
  - id: GO:0010119
    label: regulation of stomatal movement
  locations:
  - id: GO:0005886
    label: plasma membrane
  - id: GO:0005634
    label: nucleus