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First published online 11 December 2002
doi: 10.1242/jcs.00251

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Journal of Cell Science 116, 561-569 (2003)
doi: 10.1242/jcs.00251

Research Article

Nuclear oscillations and nuclear filament formation accompany single-strand annealing repair of a dicentric chromosome in Saccharomyces cerevisiae

Douglas A. Thrower*, Jennifer Stemple, Elaine Yeh and Kerry Bloom{ddagger}

Department of Biology, CB3280 University of North Carolina, Chapel Hill, NC 27599-3280, USA
* Present address: Department of Molecular, Cellular, and Developmental Biology, University of California at Santa Barbara, Santa Barbara, CA 93106, USA

{ddagger} Author for correspondence (e-mail: kerry_bloom{at}unc.edu)

Accepted 31 October 2002

Dicentric chromosomes undergo breakage during mitosis as a result of the attachment of two centromeres on one sister chromatid to opposite spindle poles. Studies utilizing a conditional dicentric chromosome III in Saccharomyces cerevisiae have shown that dicentric chromosome repair occurs primarily by deletion of one centromere via a RAD52-dependent recombination pathway. We report that dicentric chromosome resolution requires RAD1, a gene involved in the single-strand annealing DNA repair pathway. We additionally show that single-strand annealing repair of a dicentric chromosome can occur in the absence of RAD52. RAD52-independent repair requires the adaptation-defective cdc5-ad allele of the yeast polo kinase and the DNA damage checkpoint gene RAD9. Dicentric chromosome breakage in cdc5-ad rad52 mutant cells is associated with a prolonged mitotic arrest, during which nuclei undergo microtubule-dependent oscillations, accompanied by dynamic changes in nuclear morphology. We further demonstrate that the frequency of spontaneous direct repeat recombination is suppressed in yeast cells treated with benomyl, a drug that perturbs microtubules. Our findings indicate that microtubule-dependent processes facilitate recombination.

Key words: Dicentric chromosome, Microtubules, Homology search, Recombination, Single-strand annealing




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