title: Filament-dependent and -independent localization modes of Drosophila non-muscle myosin II.
abstract: Myosin II assembles into force-generating filaments that drive cytokinesis and the organization of the cell cortex. Regulation of myosin II activity can occur through modulation of filament assembly and by targeting to appropriate cellular sites. Here we show, using salt-dependent solubility and a novel fluorescence resonance energy transfer assay, that assembly of the Drosophila non-muscle myosin II heavy chain, zipper, is mediated by a 90-residue region (1849-1940) of the coiled-coil tail domain. This filament assembly domain, transiently expressed in Drosophila S2 cells, does not localize to the interphase cortex or the cytokinetic cleavage furrow, whereas a 500-residue region (1350-1865) that overlaps the NH(2) terminus of the assembly domain localizes to the interphase cortex but not the cytokinetic cleavage furrow. Targeting to these two sites appears to utilize distinct localization mechanisms as the assembly domain is required for cleavage furrow recruitment of a truncated coiled-coil tail region but not targeting to the interphase cortex. These results delineate the requirements for zipper filament assembly and indicate that the ability to form filaments is necessary for targeting to the cleavage furrow but not to the interphase cortex.
authors.affiliation: Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403.
authors.forename: Su-Ling
authors.initials: SL
authors.lastname: Liu
authors.affiliation: Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403.
authors.forename: Natasha
authors.initials: N
authors.lastname: Fewkes
authors.affiliation: Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403.
authors.forename: Derek
authors.initials: D
authors.lastname: Ricketson
authors.affiliation: Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403.
authors.forename: Rhiannon R
authors.initials: RR
authors.lastname: Penkert
authors.affiliation: Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403. Electronic address: prehoda@molbio.uoregon.edu.
authors.forename: Kenneth E
authors.initials: KE
authors.lastname: Prehoda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004330:Drosophila / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005455:Fluorescent Antibody Technique; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D008969:Molecular Sequence Data; D024462:Myosin Type II / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D029721:Drosophila Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D024462:Myosin Type II