Functional analysis of the Drosophila diaphanous FH protein in early embryonic development.
The Drosophila Formin Homology (FH) protein Diaphanous has an essential role during cytokinesis. To gain insight into the function of Diaphanous during cytokinesis and explore its role in other processes, we generated embryos deficient for Diaphanous and analyzed three cell-cycle-regulated actin-mediated events during embryogenesis: formation of the metaphase furrow, cellularization and formation of the pole cells. In dia embryos, all three processes are defective. Actin filaments do not organize properly to the metaphase and cellularization furrows and the actin ring is absent from the base of the presumptive pole cells. Furthermore, plasma membrane invaginations that initiate formation of the metaphase furrow and pole cells are missing. Immunolocalization studies of wild-type embryos reveal that Diaphanous localizes to the site where the metaphase furrow is anticipated to form, to the growing tip of cellularization furrows, and to contractile rings. In addition, the dia mutant phenotype reveals a role for Diaphanous in recruitment of myosin II, anillin and Peanut to the cortical region between actin caps. Our findings thus indicate that Diaphanous has a role in actin cytoskeleton organization and is essential for many, if not all, actin-mediated events involving membrane invagination. Based on known biochemical functions of FH proteins, we propose that Diaphanous serves as a mediator between signaling molecules and actin organizers at specific phases of the cell cycle.
Afshar
K
K
Department of Biology, Center for Molecular Genetics, University of California, San Diego, La Jolla CA 92093-0634, USA.
Stuart
B
B
Wasserman
S A
SA
D000199:Actins / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D002352:Carrier Proteins / Q000235:genetics*; D002455:Cell Division / Q000235:genetics*; D002462:Cell Membrane / Q000378:metabolism; D003285:Contractile Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004330:Drosophila / Q000196:embryology* / Q000235:genetics; D029721:Drosophila Proteins; D000080865:Formins; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D019476:Insect Proteins / Q000378:metabolism; D008677:Metaphase / Q000235:genetics; D008840:Microfilament Proteins; D009154:Mutation; D009218:Myosins / Q000378:metabolism; D010641:Phenotype; D015398:Signal Transduction
D000199:Actins; D002352:Carrier Proteins; D003285:Contractile Proteins; D029721:Drosophila Proteins; D000080865:Formins; D019476:Insect Proteins; D008840:Microfilament Proteins; C095886:anillin; C106890:diaphanous protein, Drosophila; C087195:pnut protein, Drosophila; D009218:Myosins