Kirkpatrick D T, Ferguson J R, Petes T D, Symington L S
Department of Biology and Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina 27599-3280, USA.
Genetics. 2000 Dec;156(4):1549-57. doi: 10.1093/genetics/156.4.1549.
Exonuclease I was originally identified as a 5' --> 3' deoxyribonuclease present in fractionated extracts of Schizosaccharomyces pombe and Saccharomyces cerevisiae. Genetic analysis of exo1 mutants of both yeasts revealed no major defect in meiosis, suggesting that exonuclease I is unlikely to be the primary activity that processes meiosis-specific double-strand breaks (DSBs). We report here that exo1 mutants of S. cerevisiae exhibit subtle but complex defects in meiosis. Diploids containing a homozygous deletion of EXO1 show decreased spore viability associated with an increase in meiosis I nondisjunction, while intergenic recombination is reduced about twofold. Exo1p functions in the same pathway as Msh5p for intergenic recombination. The length of heteroduplex tracts within the HIS4 gene is unaffected by the exo1 mutation. These results suggest that Exo1p is unlikely to play a major role in processing DSBs to form single-stranded tails at HIS4, but instead appears to promote crossing over to ensure disjunction of homologous chromosomes. In addition, our data indicate that exonuclease I may have a minor role in the correction of large DNA mismatches that occur in heteroduplex DNA during meiotic recombination at the HIS4 locus.
核酸外切酶I最初被鉴定为一种5'→3'脱氧核糖核酸酶,存在于粟酒裂殖酵母和酿酒酵母的分级提取物中。对这两种酵母的exo1突变体进行遗传分析,结果显示减数分裂过程中没有重大缺陷,这表明核酸外切酶I不太可能是处理减数分裂特异性双链断裂(DSB)的主要活性酶。我们在此报告,酿酒酵母的exo1突变体在减数分裂中表现出细微但复杂的缺陷。含有EXO1纯合缺失的二倍体显示孢子活力下降,与减数分裂I不分离增加有关,而基因间重组减少约两倍。Exo1p在基因间重组中与Msh5p在同一途径发挥作用。HIS4基因内异源双链区域的长度不受exo1突变的影响。这些结果表明,Exo1p不太可能在处理DSB以在HIS4处形成单链尾巴方面发挥主要作用,而是似乎促进交叉以确保同源染色体的分离。此外,我们的数据表明,核酸外切酶I可能在纠正HIS4基因座减数分裂重组期间异源双链DNA中出现的大DNA错配方面发挥次要作用。