Rattray Alison, Santoyo Gustavo, Shafer Brenda, Strathern Jeffrey N
Gene Regulation and Chromosome Biology Laboratory, NCI-Frederick, FNLCR, Frederick, Maryland, United States of America.
PLoS Genet. 2015 Jan 8;11(1):e1004910. doi: 10.1371/journal.pgen.1004910. eCollection 2015 Jan.
Mutations accumulate during all stages of growth, but only germ line mutations contribute to evolution. While meiosis contributes to evolution by reassortment of parental alleles, we show here that the process itself is inherently mutagenic. We have previously shown that the DNA synthesis associated with repair of a double-strand break is about 1000-fold less accurate than S-phase synthesis. Since the process of meiosis involves many programmed DSBs, we reasoned that this repair might also be mutagenic. Indeed, in the early 1960's Magni and Von Borstel observed elevated reversion of recessive alleles during meiosis, and found that the revertants were more likely to be associated with a crossover than non-revertants, a process that they called "the meiotic effect." Here we use a forward mutation reporter (CAN1 HIS3) placed at either a meiotic recombination coldspot or hotspot near the MAT locus on Chromosome III. We find that the increased mutation rate at CAN1 (6 to 21 -fold) correlates with the underlying recombination rate at the locus. Importantly, we show that the elevated mutation rate is fully dependent upon Spo11, the protein that introduces the meiosis specific DSBs. To examine associated recombination we selected for random spores with or without a mutation in CAN1. We find that the mutations isolated this way show an increased association with recombination (crossovers, loss of crossover interference and/or increased gene conversion tracts). Polζ appears to contribute about half of the mutations induced during meiosis, but is not the only source of mutations for the meiotic effect. We see no difference in either the spectrum or distribution of mutations between mitosis and meiosis. The correlation of hotspots with elevated mutagenesis provides a mechanism for organisms to control evolution rates in a gene specific manner.
突变在生长的各个阶段都会积累,但只有生殖系突变会推动进化。虽然减数分裂通过亲本等位基因的重排促进进化,但我们在此表明,这个过程本身就具有内在的诱变作用。我们之前已经表明,与双链断裂修复相关的DNA合成准确性比S期合成低约1000倍。由于减数分裂过程涉及许多程序性双链断裂,我们推测这种修复也可能具有诱变作用。事实上,在20世纪60年代早期,马格尼和冯·博斯特尔观察到减数分裂期间隐性等位基因的回复突变率升高,并且发现回复体比非回复体更有可能与交叉相关,他们将这个过程称为“减数分裂效应”。在这里,我们使用一个正向突变报告基因(CAN1 HIS3),将其置于III号染色体上MAT位点附近的减数分裂重组冷点或热点处。我们发现CAN1处增加的突变率(6至21倍)与该位点潜在的重组率相关。重要的是,我们表明升高的突变率完全依赖于Spo11,即引入减数分裂特异性双链断裂的蛋白质。为了检查相关的重组,我们选择了有或没有CAN1突变的随机孢子。我们发现以这种方式分离出的突变与重组(交叉、交叉干扰的丧失和/或基因转换片段增加)的关联增加。Polζ似乎在减数分裂期间诱导的突变中贡献了约一半,但不是减数分裂效应的唯一突变来源。我们在有丝分裂和减数分裂之间的突变谱或分布上没有发现差异。热点与诱变作用增强之间的相关性为生物体以基因特异性方式控制进化速率提供了一种机制。