Neufing P J, Shearwin K E, Camerotto J, Egan J B
Department of Biochemistry, University of Adelaide, Australia.
Mol Microbiol. 1996 Aug;21(4):751-61. doi: 10.1046/j.1365-2958.1996.351394.x.
We have shown previously that the cII gene product of the non-lambdoid temperate bacteriophage 186 is required for the establishment of lysogeny. We show here that CII, a potential helix-turn-helix DNA-binding protein, establishes lysogeny by activating a promoter (PE) which spans the apl/cII intergenic region, upstream of the lysogenic promoter, PL. The start site of the PE transcript (+1) has been mapped by primer extension and we have identified the CII binding determinants at PE by DNase I footprinting. CII binds to inverted repeat sequences separated by two turns of the helix, with binding half-sites centred at the 38 and -58 positions of PE. Oligomerisation studies with purified CII protein indicate that a CII tetramer may be the species that binds to this site. We also show that PE is subject to direct negative feedback by the CI repressor.
我们之前已经表明,非λ样温和噬菌体186的cII基因产物是溶原性建立所必需的。我们在此表明,CII是一种潜在的螺旋-转角-螺旋DNA结合蛋白,它通过激活一个启动子(PE)来建立溶原性,该启动子跨越溶原性启动子PL上游的apl/cII基因间区域。通过引物延伸确定了PE转录本的起始位点(+1),并且我们通过DNA酶I足迹法确定了PE处的CII结合决定簇。CII与由螺旋的两圈隔开的反向重复序列结合,结合半位点位于PE的38和-58位置的中心。对纯化的CII蛋白进行的寡聚化研究表明,CII四聚体可能是结合该位点的物种。我们还表明,PE受到CI阻遏物的直接负反馈作用。