Wagstaff J E, Klapholz S, Waddell C S, Jensen L, Esposito R E
Mol Cell Biol. 1985 Dec;5(12):3532-44. doi: 10.1128/mcb.5.12.3532-3544.1985.
We used haploid yeast cells that express both the MATa and MAT alpha mating-type alleles and contain the spo13-1 mutation to characterize meiotic recombination within single, unpaired chromosomes in Rec+ and Rec- Saccharomyces cerevisiae. In Rec+ haploids, as in diploids, intrachromosomal recombination in the ribosomal DNA was detected in 2 to 6% of meiotic divisions, and most events were unequal reciprocal sister chromatid exchange (SCE). By contrast, intrachromosomal recombination between duplicated copies of the his4 locus occurred in approximately 30% of haploid meiotic divisions, a frequency much higher than that reported in diploids; only about one-half of the events were unequal reciprocal SCE. The spo11-1 mutation, which virtually eliminates meiotic exchange between homologs in diploid meiosis, reduced the frequency of intrachromosomal recombination in both the ribosomal DNA and the his4 duplication during meiosis by 10- to greater than 50-fold. This Rec- mutation affected all forms of recombination within chromosomes: unequal reciprocal SCE, reciprocal intrachromatid exchange, and gene conversion. Intrachromosomal recombination in spo11-1 haploids was restored by transformation with a plasmid containing the wild-type SPO11 gene. Mitotic intrachromosomal recombination frequencies were unaffected by spo11-1. This is the first demonstration of a gene product required for recombination between homologs as well as recombination within chromosomes during meiosis.
我们使用了同时表达 MATa 和 MATα 交配型等位基因且含有 spo13 - 1 突变的单倍体酵母细胞,来表征 Rec⁺ 和 Rec⁻ 酿酒酵母中单条未配对染色体上的减数分裂重组。在 Rec⁺ 单倍体中,如同在二倍体中一样,在 2%至 6%的减数分裂中检测到核糖体 DNA 中的染色体内重组,并且大多数事件是不等的相互姐妹染色单体交换(SCE)。相比之下,his4 基因座重复拷贝之间的染色体内重组发生在大约 30%的单倍体减数分裂中,该频率远高于二倍体中报道的频率;只有约一半的事件是不等的相互 SCE。spo11 - 1 突变在二倍体减数分裂中几乎消除了同源染色体之间的减数分裂交换,在减数分裂期间将核糖体 DNA 和 his4 重复序列中的染色体内重组频率降低了 10 至 50 倍以上。这种 Rec⁻ 突变影响了染色体内所有形式的重组:不等的相互 SCE、相互染色单体内交换和基因转换。通过用含有野生型 SPO11 基因的质粒转化,spo11 - 1 单倍体中的染色体内重组得以恢复。有丝分裂染色体内重组频率不受 spo11 - 1 的影响。这是首次证明在减数分裂期间同源染色体之间的重组以及染色体内的重组都需要一种基因产物。