酵母交配型转换的染色体重折叠模型。

Chromosome-refolding model of mating-type switching in yeast.

作者信息

Avşaroğlu Barış, Bronk Gabriel, Li Kevin, Haber James E, Kondev Jane

机构信息

Department of Physics, Brandeis University, Waltham, MA 02454.

Department of Biology, Brandeis University, Waltham, MA 02454;

出版信息

Proc Natl Acad Sci U S A. 2016 Nov 8;113(45):E6929-E6938. doi: 10.1073/pnas.1607103113. Epub 2016 Oct 24.

Abstract

Chromosomes are folded into cells in a nonrandom fashion, with particular genetic loci occupying distinct spatial regions. This observation raises the question of whether the spatial organization of a chromosome governs its functions, such as recombination or transcription. We consider this general question in the specific context of mating-type switching in budding yeast, which is a model system for homologous recombination. Mating-type switching is induced by a DNA double-strand break (DSB) at the locus on chromosome III, followed by homologous recombination between the cut locus and one of two donor loci (α and a), located on the same chromosome. Previous studies have suggested that in a cells after the DSB is induced chromosome III undergoes refolding, which directs the locus to recombine with α. Here, we propose a quantitative model of mating-type switching predicated on the assumption of DSB-induced chromosome refolding, which also takes into account the previously measured stochastic dynamics and polymer nature of yeast chromosomes. Using quantitative fluorescence microscopy, we measure changes in the distance between the donor (α) and loci after the DSB and find agreement with the theory. Predictions of the theory also agree with measurements of changes in the use of α as the donor, when we perturb the refolding of chromosome III. These results establish refolding of yeast chromosome III as a key driving force in switching and provide an example of a cell regulating the spatial organization of its chromosome so as to direct homology search during recombination.

摘要

染色体以非随机方式折叠在细胞中,特定的基因座占据不同的空间区域。这一观察结果引发了一个问题,即染色体的空间组织是否支配其功能,如重组或转录。我们在芽殖酵母交配型转换的特定背景下考虑这个一般性问题,芽殖酵母是同源重组的一个模型系统。交配型转换由位于第三条染色体上的位点处的DNA双链断裂(DSB)诱导,随后在切割的位点与位于同一条染色体上的两个供体位点(α和a)之一之间进行同源重组。先前的研究表明,在诱导DSB后,a细胞中的第三条染色体发生重新折叠,这引导位点与α重组。在这里,我们基于DSB诱导染色体重新折叠的假设提出了一个交配型转换的定量模型,该模型还考虑了先前测量的酵母染色体的随机动力学和聚合物性质。使用定量荧光显微镜,我们测量了DSB后供体(α)和位点之间距离的变化,并发现与理论一致。当我们干扰第三条染色体的重新折叠时,该理论的预测也与α作为供体的使用变化的测量结果一致。这些结果确立了酵母第三条染色体的重新折叠是位点转换的关键驱动力,并提供了一个细胞调节其染色体空间组织以在重组过程中指导同源搜索的例子。

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