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PMID: 16292343 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The regulation of mDia1 by autoinhibition and its release by Rho*GTP.

The EMBO journal ·Vol. 24 ·No. 23 ·2005-12-07 ·Pages 4176-87

Lammers M, Rose R, Scrima A, Wittinghofer A

Abstract

Formins induce the nucleation and polymerisation of unbranched actin filaments via the formin-homology domains 1 and 2. Diaphanous-related formins (Drfs) are regulated by a RhoGTPase-binding domain situated in the amino-terminal (N-terminal) region and a carboxy-terminal Diaphanous-autoregulatory domain (DAD), whose interaction stabilises an autoinhibited inactive conformation. Binding of active Rho releases DAD and activates the catalytic activity of mDia. Here, we report on the interaction of DAD with the regulatory N-terminus of mDia1 (mDia(N)) and its release by Rho*GTP. We have defined the elements required for tight binding and solved the three-dimensional structure of a complex between an mDia(N) construct and DAD by X-ray crystallography. The core DAD region is an alpha-helical peptide, which binds in the most highly conserved region of mDia(N) using mainly hydrophobic interactions. The structure suggests a two-step mechanism for release of autoinhibition whereby Rho*GTP, although having a partially nonoverlapping binding site, displaces DAD by ionic repulsion and steric clashes. We show that Rho*GTP accelerates the dissociation of DAD from the mDia(N)*DAD complex.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Carrier Proteins/antagonists & inhibitors,genetics,metabolism Crystallography, X-Ray DNA Mutational Analysis Formins Guanosine Triphosphate/physiology Mice Molecular Sequence Data Mutation Protein Structure, Tertiary rho GTP-Binding Proteins/physiology
Chemicals
Carrier Proteins Diap1 protein, mouse Formins Guanosine Triphosphate rho GTP-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lammers Michael
Department of Structural Biology, Max-Planck-Institute of Molecular Physiology, Dortmund, Germany.
Rose Rolf
Scrima Andrea
Wittinghofer Alfred
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600879
Published
2005-12-07
Epub
2005-00-17
Pages
4176-87
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1356318
Subset
IM
Databases
PDB
2BAP
External Links
PubMed source DOI: 10.1038/sj.emboj.7600879 PMC Full Text View journal details
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