Hernández-Arriaga Ana M, Rubio-Lepe Tania S, Espinosa Manuel, del Solar Gloria
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Nucleic Acids Res. 2009 Aug;37(14):4799-811. doi: 10.1093/nar/gkp503. Epub 2009 Jun 11.
Replication of the promiscuous plasmid pMV158 requires expression of the initiator repB gene, which is controlled by the repressor CopG. Genes repB and copG are co-transcribed from promoter P(cr). We have studied the interactions between RNA polymerase, CopG and the promoter to elucidate the mechanism of repression by CopG. Complexes formed at 0 degrees C and at 37 degrees C between RNA polymerase and P(cr) differed from each other in stability and in the extent of the DNA contacted. The 37 degrees C complex was very stable (half-life of about 3 h), and shared features with typical open complexes generated at a variety of promoters. CopG protein repressed transcription from P(cr) at two different stages in the process leading to the initiation complex. First, CopG hindered binding of RNA polymerase to the promoter. Second, CopG was able to displace RNA polymerase once the enzyme has formed a stable complex with P(cr). A model for the CopG-mediated disassembly of the stable RNA polymerase-P(cr) promoter complex is presented.
杂乱质粒pMV158的复制需要起始子repB基因的表达,该基因受阻遏物CopG的控制。repB基因和copG基因从启动子P(cr)共同转录。我们研究了RNA聚合酶、CopG与启动子之间的相互作用,以阐明CopG的阻遏机制。RNA聚合酶与P(cr)在0℃和37℃形成的复合物在稳定性和所接触的DNA范围上彼此不同。37℃的复合物非常稳定(半衰期约为3小时),并具有在各种启动子处产生的典型开放复合物的特征。CopG蛋白在导致起始复合物形成的过程中的两个不同阶段抑制从P(cr)的转录。首先,CopG阻碍RNA聚合酶与启动子的结合。其次,一旦酶与P(cr)形成稳定复合物,CopG能够取代RNA聚合酶。本文提出了一个CopG介导的稳定RNA聚合酶-P(cr)启动子复合物解体的模型。