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Cited by 8Protein Kinase Regulation and GTPase SignalingCellular transport and secretionCellular Mechanics and Interactions

Divergence of Rho residue 43 impacts GEF activity

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Sloan, Christina M., Quinn, Clancy V., Peters, Justin P., Farley, Janean, Goetzinger, Chris, Wernli, Molly, DeMali, Kris A., Ellerbroek, Shawn M.·2012·Small GTPases
DOI 10.4161/sgtp.19557PMC PMC3398911
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Abstract

RhoA, RhoB and RhoC GTPases are over 85% identical at the amino acid level, with RhoA and RhoC differing at only one residue (43) across the initial two-thirds of their sequences. A putative regulatory distinction between the molecules is their capacity to be uniquely activated by guanine nucleotide exchange factors (GEFs). We hypothesize that variation of amino acid residue 43 between RhoA/B (valine) and RhoC (isoleucine) impacts GEF activity. Direct participation of residue 43 in GEF-catalyzed exchange was confirmed by the observation that mutation of this position to a threonine reduced GEF-catalyzed nucleotide exchange activity in vitro (Vav2, XPLN, GEFT, Dbl and Dbs) and greatly depressed RhoA and RhoC GTP-loading profiles in cell lysates. Using a residue swap approach, substitution of RhoA Val 43 with an Ile was found to significantly promote basal nucleotide exchange activity and enhance GTP-loading in cells. Substitution of Val 43 with an Ile in RhoB negatively affected nucleotide exchange in vitro. Substitution of RhoC Ile 43 with a Val increased GEF-catalyzed exchange in vitro. In addition, RhoC-I43V was more efficacious at driving ovarian cancer cell invasion through matrigrel than wild-type RhoC, RhoC-I43T, wild-type RhoA, RhoA-V43I or RhoA-V43T GTPases. These findings suggest that a divergence between RhoA/B and RhoC at residue 43 impacts basal and GEF-stimulated nucleotide exchange activity.

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  • XPLN shows activity toward RhoA and RhoB but not RhoC due to an amino acid difference (Ile43 vs. Val43) at the N-terminus of the Rho isoforms.

    “One example is XPLN, which shows activity toward RhoA and RhoB but not RhoC due to an amino acid difference (Ile43 vs. Val43) at the N-terminus of the Rho isoforms [20,21].”

    From RhoA Signaling in Neurodegenerative Diseases (2022) · cites this paper as [21]

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Cited by

Sorted by citations

Authors
Sloan, C. M., Quinn, C. V., Peters, J. P., Farley, J., Goetzinger, C., Wernli, M., DeMali, K. A., & Ellerbroek, S. M.
Affiliations
  • Wartburg College (US)
  • University of Iowa (US)
Corresponding author
Ellerbroek, Shawn M.
ORCID
Venue
Small GTPases
Publisher
Informa UK Limited
Published
2012-01
Volume
3
Issue
1
Pages
15-22
ISSN
  • 2154-1248 (ISSN-L)
  • 2154-1256
DOI
10.4161/sgtp.19557
PMC
PMC3398911
OpenAlex
W2002332022
Cited by
8
References
0
Citations by year
2022: 2 · 2021: 1 · 2018: 1 · 2017: 1 · 2016: 1 · 2014: 1 · 2013: 1
FWCI
0.26
Type
journal article
Language
en
Open access
Licence
cc-by-nc
Locations
  • Small GTPases (publishedVersion, cc-by-nc)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
Funders
Vedecká Grantová Agentúra MŠVVaŠ SR a SAV
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
Keywords
RhoA, RhoB, RhoC, Rho GTPases, guanine nucleotide exchange factor, Vav2, Group Embedded Figures Test, Dbl, DBS, GTP loading, Matrigel
MeSH terms
rhoC GTP-Binding Protein, Animals, Guanosine Triphosphate, Humans, Recombinant Fusion Proteins, Point Mutation, Guanine Nucleotide Exchange Factors, rho GTP-Binding Proteins, rhoA GTP-Binding Protein, rhoB GTP-Binding Protein, NIH 3T3 Cells, Cell Line, Tumor, Mice

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