Krasilnikova M M, Samadashwily G M, Krasilnikov A S, Mirkin S M
Department of Molecular Genetics, University of Illinois at Chicago, Chicago, IL 60607, USA.
EMBO J. 1998 Sep 1;17(17):5095-102. doi: 10.1093/emboj/17.17.5095.
The influence of d(G)n.d(C)n repeats on plasmid replication in Escherichia coli cells was analyzed using electrophoretic analysis of replication intermediates. These repeats impeded the replication fork in a length- and orientation-dependent manner. Unexpectedly, the replication arrest relied primarily on the repeats' transcription. When the d(C)n sequence served as the transcriptional template, both transcription and replication were blocked. This was true for transcription driven by either bacterial or phage RNA polymerases. We hypothesize that the replication fork halts after it encounters a stalled ternary complex of the RNA polymerase, the DNA template and the r(G)n transcript. This constitutes a novel mechanism for the regulation of replication elongation. The effects of this mechanism on repeat length polymorphism and genome rearrangements are discussed.
利用复制中间体的电泳分析,研究了d(G)n.d(C)n重复序列对大肠杆菌细胞中质粒复制的影响。这些重复序列以长度和方向依赖的方式阻碍复制叉。出乎意料的是,复制停滞主要依赖于重复序列的转录。当d(C)n序列作为转录模板时,转录和复制均被阻断。无论是由细菌还是噬菌体RNA聚合酶驱动的转录都是如此。我们推测,复制叉在遇到RNA聚合酶、DNA模板和r(G)n转录本的停滞三元复合物后会停止。这构成了一种调节复制延伸的新机制。讨论了该机制对重复长度多态性和基因组重排的影响。