Hayes Sidney, Asai Kengo, Chu Audrey M, Hayes Connie
Department of Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada.
Genetics. 2005 Aug;170(4):1485-99. doi: 10.1534/genetics.105.042341. Epub 2005 Jun 14.
We examined the requirement of lambda recombination functions for marker rescue of cryptic prophage genes within the Escherichia coli chromosome. We infected lysogenic host cells with lambdaimm434 phages and selected for recombinant immlambda phages that had exchanged the imm434 region of the infecting phage for the heterologous 2.6-kb immlambda region from the prophage. Phage-encoded activity, provided by either Red or NinR functions, was required for the substitution. Red(-) phages with DeltaNinR, internal NinR deletions of rap-ninH, or orf-ninC were 117-, 12-, and 5-fold reduced for immlambda rescue in a Rec(+) host, suggesting the participation of several NinR activities. RecA was essential for NinR-dependent immlambda rescue, but had slight influence on Red-dependent rescue. The host recombination activities RecBCD, RecJ, and RecQ participated in NinR-dependent recombination while they served to inhibit Red-mediated immlambda rescue. The opposite effects of several host functions toward NinR- and Red-dependent immlambda rescue explains why the independent pathways were not additive in a Rec(+) host and why the NinR-dependent pathway appeared dominant. We measured the influence of the host recombination functions and DnaB on the appearance of orilambda-dependent replication initiation and whether orilambda replication initiation was required for immlambda marker rescue.
我们研究了λ重组功能对于大肠杆菌染色体中隐蔽原噬菌体基因标记拯救的必要性。我们用λimm434噬菌体感染溶原性宿主细胞,并筛选出重组的immλ噬菌体,这些噬菌体已将感染噬菌体的imm434区域替换为来自原噬菌体的异源2.6 kb immλ区域。这种替换需要由Red或NinR功能提供的噬菌体编码活性。在Rec(+)宿主中,带有ΔNinR的Red(-)噬菌体、rap-ninH或orf-ninC的内部NinR缺失体的immλ拯救效率分别降低了117倍、12倍和5倍,这表明有几种NinR活性参与其中。RecA对于依赖NinR的immλ拯救至关重要,但对依赖Red的拯救影响较小。宿主重组活性RecBCD、RecJ和RecQ参与了依赖NinR的重组,而它们却抑制了Red介导的immλ拯救。几种宿主功能对依赖NinR和Red的immλ拯救产生相反的影响,这解释了为什么在Rec(+)宿主中这两条独立途径不是相加的,以及为什么依赖NinR的途径似乎占主导地位。我们测量了宿主重组功能和DnaB对oriλ依赖性复制起始出现的影响,以及oriλ复制起始是否是immλ标记拯救所必需的。