title: The mouse Formin mDia1 is a potent actin nucleation factor regulated by autoinhibition.
abstract: Formin proteins are widely expressed in eukaryotes and play essential roles in assembling specific cellular actin-based structures. Formins are defined by a Formin Homology 2 (FH2) domain, as well as a proline-rich FH1 domain that binds the actin monomer binding protein, profilin, and other ligands. Constructs including FH2 of budding yeast Bni1 or fission yeast Cdc12 formins nucleate actin filaments in vitro. In this study, we demonstrate that FH2-containing constructs of murine mDia1 (also called p140 mDia or Drf1) are much more potent actin nucleators than the yeast formins. FH1 is necessary for nucleation when actin monomers are profilin bound. mDia1 is a member of the Diaphanous formin subfamily (Dia), whose members contain an N-terminal Rho GTPase binding domain (GBD) and a C-terminal Diaphanous autoinhibitory domain (DAD, ). Based on cellular and in vitro binding studies, an autoinhibitory model for Dia formin regulation proposes that GBD binding to DAD inhibits Dia-induced actin remodeling, whereas Rho binding activates by releasing GBD from DAD. Supporting this model, our results show that an N-terminal mDia1 construct strongly inhibits actin nucleation by the C terminus. RhoA partially relieves inhibition but does so when bound to either GDP or GTP analogs. Both N- and C-terminal mDia1 constructs appear to be multimeric.
authors.affiliation: Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.forename: Henry N
authors.initials: HN
authors.lastname: Higgs
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002850:Chromatography, Gel; D003285:Contractile Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D000080865:Formins; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D008958:Models, Molecular; D051304:Profilins; D011485:Protein Binding / Q000502:physiology; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D000199:Actins; D002352:Carrier Proteins; D003285:Contractile Proteins; C491464:Diap1 protein, mouse; D000080865:Formins; D008840:Microfilament Proteins; C495739:Pfn1 protein, mouse; D051304:Profilins; D020741:rho GTP-Binding Proteins