Cho N Y, Choi M, Rothman-Denes L B
Department of Molecular Genetics and Cell Biology, University of Chicago, IL 60637.
J Mol Biol. 1995 Mar 3;246(4):461-71. doi: 10.1006/jmbi.1994.0098.
Transcription of the 72kb linear double-stranded DNA genome of coliphage N4 is carried out by the sequential activity of three different RNA polymerases. Early and middle viral transcripts are synthesized by two phage-coded RNA polymerases while late transcription is carried out by the Escherichia coli sigma 70-RNA polymerase. We have determined the sequences and sites of initiation of several N4 late transcripts; N4 late promoters share weak homology with the E. coli sigma 70 promoter consensus sequence. Indeed, N4 late promoters are weak templates for the host enzyme. We present evidence that the phage-coded, single-stranded DNA-binding protein (N4SSB), a protein that is required for phage DNA replication and recombination and does not bind with sequence specificity to DNA, is the activator of E. coli RNA polymerase at late N4 promoters. Models for the mechanism of action of N4SSB as a transcriptional activator are discussed.
大肠杆菌噬菌体N4的72kb线性双链DNA基因组的转录由三种不同RNA聚合酶的顺序活性完成。早期和中期病毒转录本由两种噬菌体编码的RNA聚合酶合成,而晚期转录由大肠杆菌σ70-RNA聚合酶进行。我们已经确定了几个N4晚期转录本的序列和起始位点;N4晚期启动子与大肠杆菌σ70启动子共有序列具有弱同源性。实际上,N4晚期启动子是宿主酶的弱模板。我们提供的证据表明,噬菌体编码的单链DNA结合蛋白(N4SSB),一种噬菌体DNA复制和重组所必需且不与DNA序列特异性结合的蛋白,是N4晚期启动子处大肠杆菌RNA聚合酶的激活剂。讨论了N4SSB作为转录激活剂的作用机制模型。