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
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    • "The methanol was replaced with fresh Merck ARISTAR methanol four times to remove any remaining formaldehyde or heptane and mixed after each replacement. Fixed embryos were stored in methanol at ÿ20° before processing for immunofluorescent detection as described (Therkauf and Heck 1999; Rothwell and Sullivan 2000). Developmental staging of embryos was assessed according to the numbering scheme introduced by Mary Bownes (Bownes 1975, 1982) and later refined by Eric Wieschaus and Christiane Nüsslein-Volhard (Wieschaus and Nüsslein-Volhard 1998). "
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    ABSTRACT: The condensin complex has been implicated in the higher-order organization of mitotic chromosomes in a host of model eukaryotes from yeasts to flies and vertebrates. Although chromosomes paradoxically appear to condense in condensin mutants, chromatids are not properly resolved, resulting in chromosome segregation defects during anaphase. We have examined the role of different condensin complex components in interphase chromatin function by examining the effects of various condensin mutations on position-effect variegation in Drosophila melanogaster. Surprisingly, most mutations affecting condensin proteins were often found to result in strong enhancement of variegation in contrast to what might be expected for proteins believed to compact the genome. This suggests either that the role of condensin proteins in interphase differs from their expected role in mitosis or that the way we envision condensin's activity needs to be modified to accommodate alternative possibilities.
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    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.
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    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. DOI:10.1038/35050579 · 20.06 Impact Factor
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