McBeth D L, Taylor A L
J Virol. 1983 Oct;48(1):149-56. doi: 10.1128/JVI.48.1.149-156.1983.
We examined several aspects of bacteriophage Mu development in Escherichia coli strains that carry mutations in the polA structural gene for DNA polymerase I (PolI). We found that polA mutants were markedly less efficient than PolI wild-type (PolI+) strains in their capacity to form stable Mu lysogens and to support normal lytic growth of phage Mu. The frequency of lysogenization was determined for polA mutants and their isogenic PolI+ derivatives, with the result that mutants were lysogenized 3 to 8 times less frequently than were PolI+ cells. In one-step growth experiments, we found that phage Mu grew less efficiently in polA cells than in PolI+ cells, as evidenced by a 50 to 100% increase in the latent period and a 20 to 40% decrease in mean burst size in mutant cells. A further difference noted in infected polA strains was a 10-fold reduction in the frequency of Mu-mediated transposition of chromosomal genes to an F plasmid. Pulse labeling and DNA-DNA hybridization assays to measure the rate of phage Mu DNA synthesis after the induction of thermosensitive prophages indicated that phage Mu replication began at about the same time in both polA and PolI+ strains, but proceeded at a slower rate in polA cells. We conclude that PolI is normally involved in the replication and integration of phage Mu. However, since phage Mu does not exhibit an absolute requirement for normal levels of PolI, it appears that residual PolI activity in the mutant strains, other cellular enzymes, or both can partially compensate for the absence of normal PolI activity.
我们研究了携带DNA聚合酶I(PolI)的polA结构基因突变的大肠杆菌菌株中噬菌体Mu发育的几个方面。我们发现,与PolI野生型(PolI+)菌株相比,polA突变体在形成稳定的Mu溶原菌以及支持噬菌体Mu正常裂解生长的能力上明显较低效。测定了polA突变体及其同基因PolI+衍生物的溶原化频率,结果是突变体被溶原化的频率比PolI+细胞低3至8倍。在一步生长实验中,我们发现噬菌体Mu在polA细胞中的生长效率低于在PolI+细胞中的生长效率,突变细胞中的潜伏期增加50%至100%以及平均裂解量减少20%至40%就证明了这一点。在感染的polA菌株中还注意到的另一个差异是,Mu介导的染色体基因转座到F质粒的频率降低了10倍。对温度敏感原噬菌体诱导后测量噬菌体Mu DNA合成速率的脉冲标记和DNA-DNA杂交分析表明,噬菌体Mu复制在polA和PolI+菌株中大约在同一时间开始,但在polA细胞中进行得较慢。我们得出结论,PolI通常参与噬菌体Mu的复制和整合。然而,由于噬菌体Mu对正常水平的PolI没有绝对需求,看来突变菌株中的残余PolI活性、其他细胞酶或两者都可以部分补偿正常PolI活性的缺失。