Reed M R, Shearwin K E, Pell L M, Egan J B
Biochemistry Department, University of Adelaide, South Australia.
Mol Microbiol. 1997 Feb;23(4):669-81. doi: 10.1046/j.1365-2958.1997.2521620.x.
In the present study we show that the Apl protein of the temperate coliphage 186 combines, in one protein, the activities of the coliphage lambda proteins Cro and Xis. We have shown previously that Apl represses both the lysogenic promoter, pL, and the major lytic promoter, pR, and is required for excision of the prophage. Apl binds at two locations on the phage chromosome, i.e. between pR and pL and at the phage-attachment site. Using an in vivo recombination assay, we now show that the role of Apl in excision is in the process itself and is not simply a consequence of repression of pR or pL. To study the repressive role of Apl at the switch promoters we isolated Apl-resistant operator mutants and used them to demonstrate a requirement for Apl in the efficient derepression of the lysogenic promoter during prophage induction. We conclude that Apl is both an excisionase and transcriptional repressor.
在本研究中,我们表明温和型大肠杆菌噬菌体186的Apl蛋白在一种蛋白质中兼具大肠杆菌噬菌体λ蛋白Cro和Xis的活性。我们先前已表明,Apl既能抑制溶原性启动子pL,也能抑制主要的裂解启动子pR,并且是前噬菌体切除所必需的。Apl结合在噬菌体染色体上的两个位置,即在pR和pL之间以及噬菌体附着位点。使用体内重组试验,我们现在表明Apl在切除过程中的作用在于切除过程本身,而不仅仅是抑制pR或pL的结果。为了研究Apl在开关启动子处的抑制作用,我们分离出了对Apl有抗性的操纵子突变体,并利用它们证明了在原噬菌体诱导过程中,Apl对于溶原性启动子的有效去抑制是必需的。我们得出结论,Apl既是一种切除酶,也是一种转录抑制因子。