Bird Jeremy G, Strobel Eric J, Roberts Jeffrey W
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA
Nucleic Acids Res. 2016 Aug 19;44(14):6732-40. doi: 10.1093/nar/gkw285. Epub 2016 Apr 20.
The Escherichia coli σ70 initiation factor is required for a post-initiation, promoter-proximal pause essential for regulation of lambdoid phage late gene expression; potentially, σ70 acts at other sites during transcription elongation as well. The pause is induced by σ70 binding to a repeat of the promoter -10 sequence. After σ70 binding, further RNA synthesis occurs as DNA is drawn (or 'scrunched') into the enzyme complex, presumably exactly as occurs during initial synthesis from the promoter; this synthesis then pauses at a defined site several nucleotides downstream from the active center position when σ70 first engages the -10 sequence repeat. We show that the actual pause site in the stabilized, scrunched complex is the 'elemental pause sequence' recognized from its frequent occurrence in the E. coli genome. σ70 binding and the elemental pause sequence together, but neither alone, produce a substantial transcription pause.
大肠杆菌σ70起始因子是起始后、启动子近端暂停所必需的,而这种暂停对于λ样噬菌体晚期基因表达的调控至关重要;σ70可能在转录延伸过程中的其他位点也发挥作用。这种暂停是由σ70与启动子-10序列的重复序列结合诱导的。在σ70结合后,随着DNA被拉入(或“压缩”)酶复合物中,会发生进一步的RNA合成,推测这与从启动子开始的初始合成过程完全相同;当σ70首次与-10序列重复序列结合时,这种合成随后会在活性中心位置下游几个核苷酸处的一个确定位点暂停。我们表明,在稳定的、压缩的复合物中的实际暂停位点是从其在大肠杆菌基因组中频繁出现而识别出的“基本暂停序列”。σ70结合和基本暂停序列共同作用,但单独任何一个都不会产生显著的转录暂停。