Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2021
2021
Historique:
received:
11
02
2021
accepted:
01
05
2021
entrez:
17
5
2021
pubmed:
18
5
2021
medline:
21
10
2021
Statut:
epublish
Résumé
ARHGAP36 is an atypical Rho GTPase-activating protein (GAP) family member that drives both spinal cord development and tumorigenesis, acting in part through an N-terminal motif that suppresses protein kinase A and activates Gli transcription factors. ARHGAP36 also contains isoform-specific N-terminal sequences, a central GAP-like module, and a unique C-terminal domain, and the functions of these regions remain unknown. Here we have mapped the ARHGAP36 structure-activity landscape using a deep sequencing-based mutagenesis screen and truncation mutant analyses. Using this approach, we have discovered several residues in the GAP homology domain that are essential for Gli activation and a role for the C-terminal domain in counteracting an N-terminal autoinhibitory motif that is present in certain ARHGAP36 isoforms. In addition, each of these sites modulates ARHGAP36 recruitment to the plasma membrane or primary cilium. Through comparative proteomics, we also have identified proteins that preferentially interact with active ARHGAP36, and we demonstrate that one binding partner, prolyl oligopeptidase-like protein, is a novel ARHGAP36 antagonist. Our work reveals multiple modes of ARHGAP36 regulation and establishes an experimental framework that can be applied towards other signaling proteins.
Identifiants
pubmed: 33999959
doi: 10.1371/journal.pone.0251684
pii: PONE-D-21-04769
pmc: PMC8128262
doi:
Substances chimiques
ARHGAP36 protein, human
0
GTPase-Activating Proteins
0
Protein Isoforms
0
Banques de données
Dryad
['10.5061/dryad.dz08kprv9']
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0251684Subventions
Organisme : NCI NIH HHS
ID : T32 CA009302
Pays : United States
Organisme : NCRR NIH HHS
ID : S10 RR027431
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA124435
Pays : United States
Organisme : NCRR NIH HHS
ID : S10 RR025518
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM127030
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM120007
Pays : United States
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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