Frazier M W, Mosig G
Department of Molecular Biology, Vanderbilt University, Nashville, TN 37235.
Gene. 1990 Mar 30;88(1):7-14. doi: 10.1016/0378-1119(90)90053-t.
In an Escherichia coli rpoH mutant (affecting sigma 32, heat-shock sigma factor) infected at high temperatures with wild-type T4 phage, late T4 transcription and consequently progeny production are dramatically impaired. This defect is due, in part, to insufficient activity of sigma 70 [Frazier and Mosig, J. Bacteriol. 170 (1988) 1384-1388], which is necessary to initiate early T4 transcription. Unexpectedly, however, we found that, in this rpoH host, late T4 transcription is also impaired when the temperature is raised from 30 to 42 degrees C late after infection, when T4 transcription is directed by the T4-encoded sigma factor, sigma gp55. Here, we show that a T4 gene that we call mrh (modulates rpoH), located at 14 kb on the T4 map, is responsible for the inhibition of late T4 transcription in the rpoH mutant host. T4 deletion mutants that lack the mrh gene can produce progeny in the rpoH host, but the Mrh protein, provided in trans from a plasmid-borne mrh gene, inhibits this growth. We have cloned and sequenced this T4 gene and synthesized the Mrh protein in a T7 RNA polymerase-dependent expression system. The Mr of the Mrh protein deduced from the nucleotide sequence is 13419. Gene mrh is cotranscribed with several other, yet unidentified genes, both from an early promoter downstream from the late soc gene (encoding the small outer capsid protein) and from the late soc promoter further upstream.(ABSTRACT TRUNCATED AT 250 WORDS)
在高温下被野生型T4噬菌体感染的大肠杆菌rpoH突变体(影响σ32,即热休克σ因子)中,T4晚期转录以及子代产生显著受损。这种缺陷部分归因于σ70活性不足[弗雷泽和莫西格,《细菌学杂志》170 (1988) 1384 - 1388],σ70是启动T4早期转录所必需的。然而,出乎意料的是,我们发现,在这种rpoH宿主中,当感染后期温度从30℃升高到42℃时,T4晚期转录也会受损,此时T4转录由T4编码的σ因子σgp55指导。在此,我们表明,T4图谱上位于14 kb处的一个我们称为mrh(调节rpoH)的基因,是rpoH突变宿主中T4晚期转录受到抑制的原因。缺乏mrh基因的T4缺失突变体能够在rpoH宿主中产生子代,但从质粒携带的mrh基因反式提供的Mrh蛋白会抑制这种生长。我们克隆并测序了这个T4基因,并在T7 RNA聚合酶依赖性表达系统中合成了Mrh蛋白。从核苷酸序列推导的Mrh蛋白的分子量为13419。基因mrh与其他几个尚未鉴定的基因共转录,既从晚期soc基因(编码小外 capsid 蛋白)下游的早期启动子转录,也从更上游的晚期soc启动子转录。(摘要截短于250字)