Miller J F, Malamy M H
J Bacteriol. 1984 Oct;160(1):192-8. doi: 10.1128/jb.160.1.192-198.1984.
F factor pifC, pifA, and pifB gene expression is subject to negative regulation by the product of the pifC locus (J.F. Miller and M. H. Malamy, J. Bacteriol. 156:338-347, 1983). In this paper, we describe the properties of a new regulatory site in the pif region, pifO, which is required in cis for autoregulation of pif gene expression. Spontaneous pifO mutations were isolated that allow expression of a pifC-lacZ protein fusion in the presence of pifC product in trans. Recombination of these pifO mutations onto F'lac results in increased pifA and pifB activity. Thus, a single regulatory element, pifO, regulates pifC, pifA, and pifB expression in cis. The presence of multiple copies of a fragment from the pif region carrying wild-type pifO sequences (F coordinates 42.9 to 42.43 kilobases) in trans to F'lac results in an increase in pifA and pifB activity as measured by inhibition of T7 plating. When the pifO mutations are recombined onto a plasmid carrying pifO, the resulting recombinants are greatly decreased in the ability to increase F'lac pif expression. These results suggest that increased F'lac pifA and pifB expression caused by pifO sequences in trans is a consequence of titration of pifC product and derepression of the pif operon.
F因子的pifC、pifA和pifB基因表达受到pifC位点产物的负调控(J.F.米勒和M.H.马拉米,《细菌学杂志》156:338 - 347,1983年)。在本文中,我们描述了pif区域一个新的调控位点pifO的特性,它对于pif基因表达的自动调控是顺式作用所必需的。分离到了自发的pifO突变,这些突变使得在反式存在pifC产物的情况下,pifC - lacZ蛋白融合体能够表达。将这些pifO突变重组到F'lac上会导致pifA和pifB活性增加。因此,单一的调控元件pifO顺式调控pifC、pifA和pifB的表达。在反式存在携带野生型pifO序列的pif区域片段(F坐标42.9至42.43千碱基)的多个拷贝时,通过抑制T7平板接种来测量,会导致pifA和pifB活性增加。当将pifO突变重组到携带pifO的质粒上时,所得重组体增加F'lac pif表达的能力会大大降低。这些结果表明,反式存在的pifO序列导致F'lac pifA和pifB表达增加是pifC产物被滴定以及pif操纵子去阻遏的结果。