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Vol. 13, Issue 4, 1132-1143, April 2002
Molecular and Cell Biology Laboratory, The Salk Institute, 10010 North Torrey Pines Road, La Jolla, CA 92037
The chromosomal passenger proteins aurora-B, survivin, and inner
centromere protein (INCENP) have been implicated in coordinating chromosome segregation with cell division. This work describes the
interplay between aurora, survivin, and INCENP orthologs in the fission
yeast Schizosaccharomyces pombe and defines their roles
in regulating chromosome segregation and cytokinesis. We describe the
cloning and characterization of the aurora-related kinase gene
ark1+, demonstrating that it is an
essential gene required for sister chromatid segregation. Cells lacking
Ark1p exhibit the cut phenotype, DNA fragmentation, and
other defects in chromosome segregation. Overexpression of a
kinase-defective version of Ark1, Ark1-K147R, inhibits cytokinesis,
with cells exhibiting an elongated, multiseptate phenotype. Ark1p
interacts physically and/or genetically with the survivin and INCENP
orthologs Bir1p and Pic1p. We identified Pic1p in a two-hybrid screen
for Ark1-K147R interacting partners and went on to map domains in both
proteins that mediate their binding. Pic1p residues 925-972 are
necessary and sufficient for Ark1p binding, which occurs through the
kinase domain. As with Ark1-K147R, overexpression of Ark1p-binding
fragments of Pic1p leads to multiseptate phenotypes. We also provide
evidence that the dominant-negative effect of Ark1-K147R requires Pic1p
binding, indicating that the formation of Ark1p-Pic1p complexes is
required for the execution of cytokinesis.
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