Myosin VI stabilizes an actin network during Drosophila spermatid individualization.
Here, we demonstrate a new function of myosin VI using observations of Drosophila spermatid individualization in vivo. We find that myosin VI stabilizes a branched actin network in actin structures (cones) that mediate the separation of the syncytial spermatids. In a myosin VI mutant, the cones do not accumulate F-actin during cone movement, whereas overexpression of myosin VI leads to bigger cones with more F-actin. Myosin subfragment 1-fragment decoration demonstrated that the actin cone is made up of two regions: a dense meshwork at the front and parallel bundles at the rear. The majority of the actin filaments were oriented with their pointed ends facing in the direction of cone movement. Our data also demonstrate that myosin VI binds to the cone front using its motor domain. Fluorescence recovery after photobleach experiments using green fluorescent protein-myosin VI revealed that myosin VI remains bound to F-actin for minutes, suggesting its role is tethering, rather than transporting cargo. We hypothesize that myosin VI protects the actin cone structure either by cross-linking actin filaments or anchoring regulatory molecules at the cone front. These observations uncover a novel mechanism mediated by myosin VI for stabilizing long-lived actin structures in cells.
Noguchi
Tatsuhiko
T
Department of Biology, Washington University in St. Louis, St. Louis, MO 63130, USA. noguchi@biology2.wustl.edu
Lenartowska
Marta
M
Miller
Kathryn G
KG
D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002465:Cell Movement / Q000502:physiology; D004330:Drosophila / Q000502:physiology*; D029721:Drosophila Proteins / Q000378:metabolism*; D017930:Genes, Reporter; D008297:Male; D008854:Microscopy, Electron; D018995:Myosin Heavy Chains / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013087:Spermatids / Q000502:physiology*; D013737:Testis / Q000502:physiology*
D000199:Actins; D029721:Drosophila Proteins; D011993:Recombinant Fusion Proteins; C089785:myosin VI; D018995:Myosin Heavy Chains