Liu T, Haggård-Ljungquist E
Department of Genetics, Stockholm University, S-106 91 Stockholm, Sweden.
J Virol. 1999 Dec;73(12):9816-26. doi: 10.1128/JVI.73.12.9816-9826.1999.
Phage WPhi is a member of the nonlambdoid P2 family of temperate phages. The DNA sequence of the whole early-control region and the int and attP region of phage WPhi has been determined. The phage integration site was located at 88.6 min of the Escherichia coli K-12 map, where a 47-nucleotide sequence was found to be identical in the host and phage genomes. The WPhi Int protein belongs to the Int family of site-specific recombinases, and it seems to have the same arm binding recognition sequence as P2 Int, but the core sequence differs. The transcriptional switch contains two face-to-face promoters, Pe and Pc, and two repressors, C and Cox, controlling Pe and Pc, respectively. The early Pe promoter was found to be much stronger than the Pc promoter. Furthermore, the Pe transcript was shown to interfere with Pc transcription. By site-directed mutagenesis, the binding site of the immunity repressor was located to two direct repeats spanning the Pe promoter. A point mutation in one or the other repeat does not affect repression by C, but when it is included in both, C has no effect on the Pe promoter. The Cox repressor efficiently blocks expression from the Pc promoter, but its DNA recognition sequence was not evident. Most members of the P2 family of phages are able to function as helpers for satellite phage P4, which lacks genes encoding structural proteins and packaging and lysis functions. In this work it is shown that P4 E, known to function as an antirepressor by binding to P2 C, also turns the transcriptional switch of WPhi from the lysogenic to the lytic mode. However, in contrast to P2 Cox, WPhi Cox is unable to activate the P4 Pll promoter.
噬菌体WPhi是温和噬菌体非λ样P2家族的成员。已经确定了噬菌体WPhi整个早期控制区域以及int和attP区域的DNA序列。噬菌体整合位点位于大肠杆菌K-12图谱的88.6分钟处,在宿主和噬菌体基因组中发现一个47个核苷酸的序列是相同的。WPhi Int蛋白属于位点特异性重组酶的Int家族,它似乎具有与P2 Int相同的臂结合识别序列,但核心序列不同。转录开关包含两个面对面的启动子Pe和Pc,以及两个分别控制Pe和Pc的阻遏物C和Cox。发现早期的Pe启动子比Pc启动子强得多。此外,Pe转录本显示会干扰Pc转录。通过定点诱变,免疫阻遏物的结合位点定位于跨越Pe启动子的两个直接重复序列。一个或另一个重复序列中的点突变不影响C的阻遏作用,但当两个重复序列都包含该点突变时,C对Pe启动子没有作用。Cox阻遏物有效地阻断了Pc启动子的表达,但其DNA识别序列不明显。P2家族的大多数噬菌体能够作为卫星噬菌体P4的辅助噬菌体发挥作用,P4缺乏编码结构蛋白、包装和裂解功能的基因。在这项工作中表明,已知通过与P2 C结合而发挥抗阻遏作用的P4 E,也能将WPhi的转录开关从溶原模式转变为裂解模式。然而,与P2 Cox不同,WPhi Cox无法激活P4 Pll启动子。