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Cited by 2Protein Kinase Regulation and GTPase SignalingGenetics and Neurodevelopmental DisordersGenomics and Rare Diseases

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Genetic variants linked to neurodevelopmental disorders within the β3-β4 loop of the TRIO PH2 domain release autoinhibition of GEF2 activity

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Carrizales, Melissa G., Boulton, Andrew D., Koleske, Anthony J.·2025·Journal of Biological Chemistry
DOI 10.1016/j.jbc.2025.110429PMC PMC12309612

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Abstract

The TRIO protein contains two guanine exchange factor (GEF) domains, GEF1 and GEF2, which coordinate cytoskeletal rearrangements by activating Rho family GTPases. Rare variants that impact TRIO GEF1 function are associated with autism spectrum disorder, developmental delay, and intellectual disability, but variants are also found throughout the gene. GEF1 promotes GTP exchange on Rac1 and RhoG, while GEF2 activates RhoA. Although GEF1 and GEF2 share a common architecture, the pleckstrin homology (PH) domain in TRIO GEF1 (PH1) assists its activity, while the PH domain in GEF2 (PH2) inhibits its activation of RhoA. A series of single-point variants in the unique β3-β4 loop of TRIO PH2 has been identified in patients with neurodevelopmental disorders (NDDs), but how they impact TRIO GEF2 activity is not known. Using an in vitro fluorescence-based assay to assess GEF2 exchange activity on RhoA, we demonstrate that variants within the β3-β4 loop relieve GEF2 autoinhibition. Activation of RhoA inhibits neurite outgrowth in Neuro-2A (N2A) cells. GEF2 expression in N2A cells significantly reduces neurite outgrowth, and expression of the G2211E activating GEF2 variant enhances this effect. Together, our findings reveal key interactions and structural constraints for GEF2 autoinhibition and how this mechanism is a target for disruption by NDD-associated variations.

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Metadata

Authors
Carrizales, M. G., Boulton, A. D., & Koleske, A. J.
Affiliations
Yale University (US)
Corresponding author
Koleske, Anthony J.
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2025-08
Volume
301
Issue
8
Pages
110429
ISSN
0021-9258 (ISSN-L)
DOI
10.1016/j.jbc.2025.110429
PMC
PMC12309612
OpenAlex
W4411682348
Cited by
2
References
1
Citations by year
2026: 1 · 2025: 1
FWCI
0.60
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
National Institutes of Health (R01MH133562, F31NS131038, R01MH132685)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genomics and Rare Diseases
Keywords
trios, guanine nucleotide exchange factor, autoinhibition, RhoA activation, Rho GTPases, neurodevelopmental disorders, neurite outgrowth, autism spectrum disorder, intellectual disability, pleckstrin homology domain
MeSH terms
Animals, Humans, Protein Serine-Threonine Kinases, Guanine Nucleotide Exchange Factors, rhoA GTP-Binding Protein, Mice, HEK293 Cells, Rho Guanine Nucleotide Exchange Factors, Neurodevelopmental Disorders

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