Activated RhoA binds to the Pleckstrin Homology (PH) domain of PDZ-RhoGEF, a potential site for autoregulation

Zhe Chen, Frank Medina, Mu Ya Liu, Celestine Thomas, Stephen R. Sprang, Paul C. Sternweis

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Guanine nucleotide exchange factors (GEFs) catalyze exchange of GDP for GTP by stabilizing the nucleotide-free state of the small GTPases through their Dbl homology/pleckstrin homology (DH·PH) domains. Unconventionally, PDZ-RhoGEF (PRG), a member of the RGS-RhoGEFs, binds tightly to both nucleotide-free and activated RhoA (RhoA·GTP). We have characterized the interaction between PRG and activated RhoA and determined the structure of the PRG-DH·PH-RhoA·GTPγS (guanosine 5′-O-[γ-thio] triphosphate) complex. The interface bears striking similarity to a GTPase-effector interface and involves the switch regions in RhoA and a hydrophobic patch in PRG-PH that is conserved among all Lbc RhoGEFs. The two surfaces that bind activated and nucleotide-free RhoA on PRGD-H ·PH do not overlap, and a ternary complex of PRG-DH·PH bound to both forms of RhoA can be isolated by size-exclusion chromatography. This novel interaction between activated RhoA and PH could play a key role in regulation of RhoGEF activity in vivo.

Original languageEnglish (US)
Pages (from-to)21070-21081
Number of pages12
JournalJournal of Biological Chemistry
Volume285
Issue number27
DOIs
StatePublished - Jul 2 2010

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Rho Guanine Nucleotide Exchange Factors
Homeostasis
Nucleotides
Guanine Nucleotide Exchange Factors
Monomeric GTP-Binding Proteins
Size exclusion chromatography
Pleckstrin Homology Domains
platelet protein P47
Guanosine
GTP Phosphohydrolases
Guanosine Triphosphate
Gel Chromatography
Switches

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Activated RhoA binds to the Pleckstrin Homology (PH) domain of PDZ-RhoGEF, a potential site for autoregulation. / Chen, Zhe; Medina, Frank; Liu, Mu Ya; Thomas, Celestine; Sprang, Stephen R.; Sternweis, Paul C.

In: Journal of Biological Chemistry, Vol. 285, No. 27, 02.07.2010, p. 21070-21081.

Research output: Contribution to journalArticle

Chen, Zhe ; Medina, Frank ; Liu, Mu Ya ; Thomas, Celestine ; Sprang, Stephen R. ; Sternweis, Paul C. / Activated RhoA binds to the Pleckstrin Homology (PH) domain of PDZ-RhoGEF, a potential site for autoregulation. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 27. pp. 21070-21081.
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