Nuclear oscillations and nuclear filament formation accompany single-strand annealing repair of a dicentric chromosome in Saccharomyces cerevisiae.
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.
Thrower
Douglas A
DA
Department of Biology, CB3280 University of North Carolina, Chapel Hill, NC 27599-3280, USA.
Stemple
Jennifer
J
Yeh
Elaine
E
Bloom
Kerry
K
D001542:Benomyl / Q000494:pharmacology; D001683:Biological Clocks / Q000187:drug effects / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics; D022003:Cell Nucleus Structures / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D002503:Centromere / Q000235:genetics; D002875:Chromosomes / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics*; D045643:DNA Repair Enzymes; D004277:DNA, Single-Stranded / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D029721:Drosophila Proteins; D004720:Endonucleases / Q000235:genetics*; D005659:Fungicides, Industrial / Q000494:pharmacology; D008870:Microtubules / Q000187:drug effects / Q000235:genetics; D008938:Mitosis / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D051721:Rad52 DNA Repair and Recombination Protein; D011995:Recombination, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins
D018797:Cell Cycle Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D005659:Fungicides, Industrial; C497197:RAD52 protein, S cerevisiae; D051721:Rad52 DNA Repair and Recombination Protein; D029701:Saccharomyces cerevisiae Proteins; C069930:rad9 protein; D017346:Protein Serine-Threonine Kinases; D004720:Endonucleases; C069266:RAD1 protein, S cerevisiae; D045643:DNA Repair Enzymes; D001542:Benomyl