title: Mechanism of actin filament nucleation by the bacterial effector VopL.
abstract: Vibrio parahaemolyticus protein L (VopL) is an actin nucleation factor that induces stress fibers when injected into eukaryotic host cells. VopL contains three N-terminal Wiskott-Aldrich homology 2 (WH2) motifs and a unique VopL C-terminal domain (VCD). We describe crystallographic and biochemical analyses of filament nucleation by VopL. The WH2 element of VopL does not nucleate on its own and requires the VCD for activity. The VCD forms a U-shaped dimer in the crystal, stabilized by a terminal coiled coil. Dimerization of the WH2 motifs contributes strongly to nucleation activity, as do contacts of the VCD to actin. Our data lead to a model in which VopL stabilizes primarily lateral (short-pitch) contacts between actin monomers to create the base of a two-stranded filament. Stabilization of lateral contacts may be a common feature of actin filament nucleation by WH2-based factors.
authors.affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Bingke
authors.initials: B
authors.lastname: Yu
authors.forename: Hui-Chun
authors.initials: HC
authors.lastname: Cheng
authors.forename: Chad A
authors.initials: CA
authors.lastname: Brautigam
authors.forename: Diana R
authors.initials: DR
authors.lastname: Tomchick
authors.forename: Michael K
authors.initials: MK
authors.lastname: Rosen
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000737:chemistry / Q000378:metabolism; D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D018360:Crystallography, X-Ray; D019281:Dimerization; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D014736:Vibrio parahaemolyticus / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D000199:Actins; D001426:Bacterial Proteins; D008840:Microfilament Proteins