Requirement of the basic region of N-WASP/WAVE2 for actin-based motility.
WASP family proteins activate nucleation by the Arp2/3 complex, inducing rapid actin polymerization in vitro. Although the C-terminal portion of WASP family proteins (VCA) activates nucleation by the Arp2/3 complex in pure systems, we find that this fragment lacks activity in cell extracts. Thus, polystyrene beads coated with VCA did not move in brain cytosol, while beads coated with N-WASP or WAVE2 did move. The basic clusters between the WH1 domain and the CRIB domain of N-WASP were critical for movement since beads coated with N-WASP or WAVE2 constructs missing the basic clusters (Delta basic) also did not move. Furthermore, VCA and N-WASP/WAVE2 Delta basic constructs were much less able than wild-type N-WASP and WAVE2 to induce actin polymerization in cytosol. All of the proteins, with or without the basic domain, were potent activators of nucleation by purified Arp2/3 complex.
Suetsugu
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S
Department of Biochemistry, University of Tokyo, Tokyo, Japan.
Miki
H
H
Yamaguchi
H
H
Takenawa
T
T
D000199:Actins / Q000502:physiology*; D000818:Animals; D001921:Brain / Q000502:physiology; D002417:Cattle; D003599:Cytoskeleton / Q000737:chemistry / Q000502:physiology; D066298:In Vitro Techniques; D008840:Microfilament Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D008863:Microspheres; D008954:Models, Biological; D009068:Movement / Q000502:physiology; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal
D000199:Actins; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal