The CDC20 gene product of Saccharomyces cerevisiae, a beta-transducin homolog, is required for a subset of microtubule-dependent cellular processes.
Previous analysis of cdc20 mutants of the yeast Saccharomyces cerevisiae suggests that the CDC20 gene product (Cdc20p) is required for two microtubule-dependent processes, nuclear movements prior to anaphase and chromosome separation. Here we report that cdc20 mutants are defective for a third microtubule-mediated event, nuclear fusion during mating of G1 cells, but appear normal for a fourth microtubule-dependent process, nuclear migration after DNA replication. Therefore, Cdc20p is required for a subset of microtubule-dependent processes and functions at multiple stages in the life cycle. Consistent with this interpretation, we find that cdc20 cells arrested by alpha-factor or at the restrictive temperature accumulate anomalous microtubule structures, as detected by indirect immunofluorescence. The anomalous microtubule staining patterns are due to cdc20 because intragenic revertants that revert the temperature sensitivity have normal microtubule morphologies. cdc20 mutants have a sevenfold increase in the intensity of antitubulin fluorescence in intranuclear spindles compared with spindles from wild-type cells, yet the total amount of tubulin is indistinguishable by Western immunoblot analysis. This result suggests that Cdc20p modulates microtubule structure in wild-type cells either by promoting microtubule disassembly or by altering the surface of the microtubules. Finally, we cloned and sequenced CDC20 and show that it encodes a member of a family of proteins that share homology to the beta subunit of transducin.
Department of Biology, University of Virginia, Charlottesville 22901.
N
N
Sethi
M C
MC
Monteagudo
D
D
Koshland
E
E
Hogan
D J
DJ
Burke
D000595:Amino Acid Sequence; D001483:Base Sequence; D003433:Crosses, Genetic; D004260:DNA Repair; D004271:DNA, Fungal / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005838:Genotype; D008870:Microtubules / Q000502:physiology*; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D012689:Sequence Homology, Nucleic Acid; D015293:Transducin / Q000235:genetics*; D014404:Tubulin / Q000032:analysis / Q000235:genetics
D004271:DNA, Fungal; D005656:Fungal Proteins; D014404:Tubulin; D015293:Transducin