Henderson Kiersten A, Keeney Scott
Molecular Biology Program, Memorial Sloan-Kettering Cancer Center and Weill Graduate School of Medical Sciences, Cornell University, New York, NY 10021, USA.
Proc Natl Acad Sci U S A. 2004 Mar 30;101(13):4519-24. doi: 10.1073/pnas.0400843101. Epub 2004 Mar 18.
During meiosis, homologous chromosomes recombine and become closely apposed along their lengths within the synaptonemal complex (SC). In part because Spo11 is required both to make the double-strand breaks (DSBs) that initiate recombination and to promote normal SC formation in many organisms, it is clear that these two processes are intimately coupled. The molecular nature of this linkage is not well understood, but it has been proposed that SC formation initiates locally at the sites of ongoing recombination and in particular at the subset of sites that will eventually give rise to crossovers. To test this hypothesis, we examined further the relationship between DSBs and SC formation in Saccharomyces cerevisiae. SCs were monitored in a series of spo11 missense mutants with varying DSB frequencies. Alleles that blocked DSB formation gave SC phenotypes indistinguishable from a deletion mutant, and partial loss-of-function mutations with progressively more severe DSB defects caused corresponding defects in SC formation. These results strongly correlate SC formation with Spo11 catalytic activity per se. Numbers of Zip3 complexes on chromosomes, thought to represent the sites of SC initiation, also declined when Spo11 activity decreased, but in a markedly nonlinear fashion: hypomorphic spo11 alleles caused larger defects in DSB formation than in Zip3 complex formation. This nonlinear response of Zip3 closely paralleled the response of crossover recombination products. The quantitative relationship between Zip3 foci, SC formation, and crossing over strongly implicates crossover-designated recombination intermediates as the sites of SC initiation.
在减数分裂过程中,同源染色体发生重组,并在联会复合体(SC)中沿着它们的长度紧密并列。部分原因是,在许多生物体中,Spo11对于引发重组的双链断裂(DSB)的产生以及促进正常的SC形成都是必需的,很明显这两个过程紧密相关。这种联系的分子本质尚不清楚,但有人提出,SC形成在正在进行重组的位点局部起始,特别是在最终会产生交叉的位点子集中起始。为了验证这一假设,我们进一步研究了酿酒酵母中DSB与SC形成之间的关系。在一系列具有不同DSB频率的spo11错义突变体中监测SC。阻断DSB形成的等位基因产生的SC表型与缺失突变体无法区分,而具有逐渐更严重DSB缺陷的部分功能丧失突变导致SC形成出现相应缺陷。这些结果强烈表明SC形成与Spo11催化活性本身相关。当Spo11活性降低时,染色体上被认为代表SC起始位点的Zip3复合体数量也会减少,但呈明显的非线性方式:亚效spo11等位基因在DSB形成中造成的缺陷比在Zip3复合体形成中更大。Zip3的这种非线性反应与交叉重组产物的反应密切平行。Zip3焦点、SC形成和交叉之间的定量关系强烈表明,指定交叉的重组中间体是SC起始的位点。