Appasani K, Thaler D S, Goldberg E B
Department of Molecular Biology and Microbiology, Tufts University Medical School, Boston, Massachusetts 02111-1800, USA.
J Bacteriol. 1999 Feb;181(4):1352-5. doi: 10.1128/JB.181.4.1352-1355.1999.
The T4 head protein, gp2, promotes head-tail joining during phage morphogenesis and is also incorporated into the phage head. It protects the injected DNA from degradation by exonuclease V during the subsequent infection. In this study, we show that recombinant gp2, a very basic protein, rapidly kills the cells in which it is expressed. To further illustrate the protectiveness of gp2 for DNA termini, we compare the effect of gp2 expression on Red-mediated and Int-mediated recombination. Red-mediated recombination is nonspecific and requires the transient formation of double-stranded DNA termini. Int-mediated recombination, on the other hand, is site specific and does not require chromosomal termini. Red-mediated recombination is inhibited to a much greater extent than is Int-mediated recombination. We conclude from the results of these physiological and genetic experiments that T4 gp2 expression, like Mu Gam expression, kills bacteria by binding to double-stranded DNA termini, the most likely mode for its protection of entering phage DNA from exonuclease V.
T4噬菌体头部蛋白gp2在噬菌体形态发生过程中促进头尾连接,并且也会整合到噬菌体头部。在随后的感染过程中,它可保护注入的DNA不被核酸外切酶V降解。在本研究中,我们发现重组gp2(一种碱性很强的蛋白)能迅速杀死其表达所在的细胞。为了进一步阐明gp2对DNA末端的保护作用,我们比较了gp2表达对Red介导的重组和Int介导的重组的影响。Red介导的重组是非特异性的,需要双链DNA末端的瞬时形成。另一方面,Int介导的重组是位点特异性的,不需要染色体末端。Red介导的重组比Int介导的重组受到的抑制程度要大得多。从这些生理学和遗传学实验结果我们得出结论,T4 gp2的表达与Mu Gam的表达一样,通过与双链DNA末端结合来杀死细菌,这很可能是其保护进入的噬菌体DNA免受核酸外切酶V作用的方式。