Molecular dissection of cytokinesis by RNA interference in Drosophila cultured cells.
We have used double-stranded RNA-mediated interference (RNAi) to study Drosophila cytokinesis. We show that double-stranded RNAs for anillin, acGAP, pavarotti, rho1, pebble, spaghetti squash, syntaxin1A, and twinstar all disrupt cytokinesis in S2 tissue culture cells, causing gene-specific phenotypes. Our phenotypic analyses identify genes required for different aspects of cytokinesis, such as central spindle formation, actin accumulation at the cell equator, contractile ring assembly or disassembly, and membrane behavior. Moreover, the cytological phenotypes elicited by RNAi reveal simultaneous disruption of multiple aspects of cytokinesis. These phenotypes suggest interactions between central spindle microtubules, the actin-based contractile ring, and the plasma membrane, and lead us to propose that the central spindle and the contractile ring are interdependent structures. Finally, our results indicate that RNAi in S2 cells is a highly efficient method to detect cytokinetic genes, and predict that genome-wide studies using this method will permit identification of the majority of genes involved in Drosophila mitotic cytokinesis.
Istituto Pasteur-Fondazione Cenci Bolognetti and Centro di Genetica Evoluzionistica del Consiglio Nazionale delle Richerche, Dipartimento di Genetica e Biologia Molecolare, Università di Roma "La Sapienza," 00185 Rome, Italy.
Maria Patrizia
MP
Somma
Barbara
B
Fasulo
Giovanni
G
Cenci
Enrico
E
Cundari
Maurizio
M
Gatti
D000199:Actins / Q000378:metabolism; D000818:Animals; D002455:Cell Division / Q000235:genetics* / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D003285:Contractile Proteins / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology / Q000502:physiology*; D005434:Flow Cytometry; D018816:Genes, cdc; D007150:Immunohistochemistry; D008870:Microtubules / Q000378:metabolism; D010641:Phenotype; D034622:RNA Interference; D008941:Spindle Apparatus / Q000378:metabolism
D000199:Actins; D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins