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

Melissa G Carrizales,

AndrewJ.M. Boulton,

Anthony J. Koleske

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Апрель 14, 2025

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

Язык: Английский

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

Melissa G Carrizales,

AndrewJ.M. Boulton,

Anthony J. Koleske

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Апрель 14, 2025

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

Язык: Английский

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