Identification and characterization of mitotic mutations in Drosophila.

Department of Biochemistry and Cell Biology, State University of New York at Stony Brook 11794-5215, USA.
Methods in cell biology (Impact Factor: 1.44). 02/1999; 61:317-46.
Source: PubMed
  • [Show abstract] [Hide abstract]
    ABSTRACT: During early embryogenesis of Drosophila melanogaster, mutations in the DNA-replication checkpoint lead to chromosome-segregation failures. Here we show that these segregation failures are associated with the assembly of an anastral microtubule spindle, a mitosis-specific loss of centrosome function, and dissociation of several components of the gamma-tubulin ring complex from a core centrosomal structure. The DNA-replication inhibitor aphidicolin and DNA-damaging agents trigger identical mitotic defects in wild-type embryos, indicating that centrosome inactivation is a checkpoint-independent and mitosis-specific response to damaged or incompletely replicated DNA. We propose that centrosome inactivation is part of a damage-control system that blocks chromosome segregation when replication/damage checkpoint control fails.
    Nature Cell Biology 03/2000; 2(2):90-5. · 20.06 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Here we briefly review techniques used to flatten cells that otherwise round in culture, so that their division can be more clearly analyzed in vitro by high resolution light microscopy. We then describe an agar overlay procedure for use with isolated Drosophila neuroblasts, which promotes their long-term viability while also allowing for correlative studies of the same cell in the living and fixed state. This same procedure can also be used to obtain high temporal and spatial resolution images of mitosis and cytokinesis in cultured Drosophila Schneider S2 cells, which are a popular model for RNAi studies.
    Cell Motility and the Cytoskeleton 11/2003; 56(3):141-6. · 4.19 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In Drosophila syncytial blastoderm embryos, centrosomes specify the position of actin-based interphase caps and mitotic furrows. Mutations in the scrambled locus prevent assembly of mitotic furrows, but do not block actin cap formation. The scrambled gene encodes a protein that localizes to the mitotic furrows and centrosomes. Sced localization, actin reorganization from caps into mitotic furrows, and centrosome-coordinated assembly of actin caps are not blocked by microtubule disruption. Our results indicate that centrosomes may coordinate assembly of cortical actin caps through a microtubule-independent mechanism, and that Scrambled mediates a second microtubule-independent process that drives mitotic furrow assembly.
    Nature Cell Biology 02/2001; 3(1):68-75. · 20.06 Impact Factor