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Genetic evidence for a role of phospholipase C at the budding yeast kinetochore

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Abstract

Chromosome segregation during mitosis requires kinetochores, specialized organelles that mediate chromosome attachment to spindle microtubules. We have shown previously that in budding yeast, Plc1p (phosphoinositide-specific phospholipase C) localizes to centromeric loci, associates with the kinetochore proteins Ndc10p and Cep3p, and affects the function of kinetochores. Deletion of PLC1 results in nocodazole sensitivity, mitotic delay, and a higher frequency of chromosome loss. We report here that despite the nocodazole sensitivity of plc1Δ cells, Plc1p is not required for the spindle checkpoint. However, plc1Δ cells require a functional BUB1/BUB3-dependent spindle checkpoint for viability. PLC1 displays strong genetic interactions with genes encoding components of the inner kinetochore, including NDC10, SKP1, MIF2, CEP1, CEP3, and CTF13. Furthermore, plc1Δ cells display alterations in chromatin structure in the core centromere. Chromatin immunoprecipitation experiments indicate that Plc1p localizes to centromeric loci independently of microtubules, and accumulates at the centromeres during G2/M stage of cell cycle. These results are consistent with the view that Plc1p affects kinetochore function, possibly by modulating the structure of centromeric chromatin.

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Acknowledgements

This work was supported by grants from the National Institutes of Health (1 R15 GM62183-01) and the American Cancer Society (RSG-01-145-01-CCG) to A.V. We thank Drs. Elledge, Huffaker, Hyman, Kaplan, Koshland, Meluh, Murray, and Saunders, for strains and plasmids, and Dr. Vancurova for critical reading of the manuscript and helpful comments. The work has been carried out in compliance with current U.S. laws governing genetic experimentation

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  1. Department of Biological Sciences, St. John's University, 8000 Utopia Parkway, Jamaicam New York, NY 11439, USA

    N. DeLillo, C. Romero, H. Lin & A. Vancura

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  1. N. DeLillo
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  2. C. Romero
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  3. H. Lin
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  4. A. Vancura
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Correspondence to A. Vancura.

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Communicated by C. P. Hollenberg

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DeLillo, N., Romero, C., Lin, H. et al. Genetic evidence for a role of phospholipase C at the budding yeast kinetochore. Mol Gen Genomics 269, 261–270 (2003). https://doi.org/10.1007/s00438-003-0832-4

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