Danilova Olessia, Reyes-Lamothe Rodrigo, Pinskaya Marina, Sherratt David, Possoz Christophe
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK.
Mol Microbiol. 2007 Sep;65(6):1485-92. doi: 10.1111/j.1365-2958.2007.05881.x.
The circular Escherichia coli chromosome is organized by bidirectional replication into two equal left and right arms (replichores). Each arm occupies a separate cell half, with the origin of replication (oriC) at mid-cell. E. coli MukBEF belongs to the ubiquitous family of SMC protein complexes that play key roles in chromosome organization and processing. In mukBEF mutants, viability is restricted to low temperature with production of anucleate cells, reflecting chromosome segregation defects. We show that in mukB mutant cells, the two chromosome arms do not separate into distinct cell halves, but extend from pole to pole with the oriC region located at the old pole. Mutations in topA, encoding topoisomerase I, do not suppress the aberrant positioning of chromosomal loci in mukB cells, despite suppressing the temperature-sensitivity and production of anucleate cells. Furthermore, we show that MukB and the oriC region generally colocalize throughout the cell cycle, even when oriC localization is aberrant. We propose that MukBEF initiates the normal bidirectional organization of the chromosome from the oriC region.
环状的大肠杆菌染色体通过双向复制被组织成两个相等的左臂和右臂(复制子)。每个臂占据细胞的一个独立半区,复制起点(oriC)位于细胞中部。大肠杆菌MukBEF属于普遍存在的SMC蛋白复合体家族,该家族在染色体组织和加工过程中起关键作用。在mukBEF突变体中,生存能力仅限于低温,会产生无核细胞,这反映出染色体分离缺陷。我们发现,在mukB突变体细胞中,两条染色体臂不会分离到不同的细胞半区,而是从一极延伸到另一极,oriC区域位于旧的那一极。编码拓扑异构酶I的topA基因突变,尽管能抑制温度敏感性和无核细胞的产生,但并不能抑制mukB细胞中染色体位点的异常定位。此外,我们还发现,即使oriC定位异常,MukB和oriC区域在整个细胞周期中通常也会共定位。我们提出,MukBEF从oriC区域启动染色体的正常双向组织。