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Molecular Biology

, Volume 52, Issue 4, pp 598–603 | Cite as

Mutations in the Effector Domain of RhoV GTPase Impair Its Binding to Pak1 Protein Kinase

  • I. V. Korobko
  • M. V. Shepelev
Molecular Cell Biology
  • 20 Downloads

Abstract

Atypical RhoV GTPase (Chp/Wrch-2) is a member of the human Rho GTPase family, which belongs to the superfamily of Ras-related small GTPases. The biological functions of RhoV, regulation of its activity, and mechanisms of its action remain largely unexplored. Rho GTPases regulate a wide range of cellular processes by interacting with protein targets called effectors. Several putative RhoV effectors have been identified, including protein kinases of the Pak (p21-activated kinase) family: Pak1, Pak2, Pak4, and Pak6. RhoV GTPase activates Pak1 protein kinase and simultaneously induces its ubiquitin-dependent degradation. Pak1 regulates E-cadherin localization at adherens junctions downstream of RhoV during gastrulation in fish. The effector domain of RhoV mediates its binding to the CRIB (Cdc42/Rac1 interactive binding) motif in the N-terminal p21-binding domain (PBD) of Pak6 protein kinase. The role of the RhoV effector domain in mediating interaction with Pak1 has not been studied. This study has identified mutations in the effector domain of RhoV GTPase (Y60K, T63A, L65A, and D66A) that impair its interaction with Pak1 in the GST-PAK-PBD pull-down assay and coimmunoprecipitation. Our results suggest that the effector domain of RhoV mediates its binding to Pak1, complementing the current view of the molecular basics of RhoV binding to effectors of the Pak family. These data lay the basis for further studies on the role of Pak1 in RhoV-activated signaling pathways and cellular processes.

Keywords

Chp/RhoV GTPase Pak1 protein kinase Rho GTPases effector domain GST-PAK-PBD pull-down 

Abbreviations

PBD

p21-binding domain of protein kinase Pak1

GST

glutathion-S-transferase

coIP

coimmunoprecipitation

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Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  1. 1.Institute of Gene BiologyRussian Academy of SciencesMoscowRussia

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