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耐辐射细菌耐辐射球菌中的类核组织

Nucleoid organization in the radioresistant bacterium Deinococcus radiodurans.

作者信息

Passot Fanny Marie, Nguyen Hong Ha, Dard-Dascot Cloelia, Thermes Claude, Servant Pascale, Espéli Olivier, Sommer Suzanne

机构信息

Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris Sud, Bâtiment 409, Orsay, 91405, France.

Plateforme Intégrée IMAGIF - CNRS, Avenue de la Terrasse, Gif sur Yvette, 91198, France.

出版信息

Mol Microbiol. 2015 Aug;97(4):759-74. doi: 10.1111/mmi.13064. Epub 2015 Jun 25.

Abstract

Processes favoring the exceptional resistance to genotoxic stress of Deinococcus radiodurans are not yet completely characterized. It was postulated that its nucleoid and chromosome(s) organization could participate in the DNA double strand break repair process. Here, we investigated the organization of chromosome 1 by localization of three chromosomal loci including oriC, Ter and a locus located in its left arm. For this purpose, we used a ParB-parS system to visualize the position of the loci before and after exposure to γ-rays. By comparing the number of fluorescent foci with the number of copies of the studied loci present in the cells measured by quantitative polymerase chain reaction (qPCR), we demonstrated that the 4-10 copies of chromosome 1 per cell are dispersed within the nucleoid before irradiation, indicating that the chromosome copies are not prealigned. Chromosome segregation is progressive but not co-ordinated, allowing each locus to be paired with its sister during part of the cell cycle. After irradiation, the nucleoid organization is modified, involving a transient alignment of the loci in the late stage of DNA repair and a delay of segregation of the Ter locus. We discuss how these events can influence DNA double strand break repair.

摘要

嗜放射球菌对基因毒性应激具有超强抗性的相关机制尚未完全明确。据推测,其类核和染色体的组织形式可能参与了DNA双链断裂修复过程。在此,我们通过对包括oriC、Ter以及位于其左臂的一个位点在内的三个染色体位点进行定位,研究了1号染色体的组织形式。为此,我们使用了ParB-parS系统来观察γ射线照射前后这些位点的位置。通过将荧光焦点的数量与定量聚合酶链反应(qPCR)测得的细胞中所研究位点的拷贝数进行比较,我们证明了每个细胞中1号染色体的4 - 10个拷贝在辐照前分散于类核中,这表明染色体拷贝并未预先排列。染色体分离是渐进的,但并非协同进行,使得每个位点在细胞周期的部分时间内与其姐妹位点配对。辐照后,类核组织发生改变,包括在DNA修复后期位点的短暂排列以及Ter位点分离的延迟。我们讨论了这些事件如何影响DNA双链断裂修复。

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