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宿主DNA回旋酶参与噬菌体T5的生长。

Involvement of host DNA gyrase in growth of bacteriophage T5.

作者信息

Constantinou A, Voelkel-Meiman K, Sternglanz R, McCorquodale M M, McCorquodale D J

出版信息

J Virol. 1986 Mar;57(3):875-82. doi: 10.1128/JVI.57.3.875-882.1986.

Abstract

Bacteriophage T5 did not grow at the nonpermissive temperature of 42 degrees C in Escherichia coli carrying a temperature-sensitive mutation in gyrB [gyrB(Ts)], but it did grow in gyrA(Ts) mutants at 42 degrees C. These findings indicate that the A subunit of host DNA gyrase is unnecessary, whereas the B subunit is necessary for growth of T5. The necessity for the B subunit was confirmed by a strong inhibition of T5 growth by novobiocin and coumermycin A1, which interfere specifically with the function of the B subunit of host DNA gyrase. However, T5 growth was also strongly inhibited by nalidixic acid, which interferes specifically with the function of the A subunit. This inhibition was due to the interaction of nalidixic acid with the A subunit and not just to its binding to DNA, because appropriate mutations in the gyrA gene of the host conferred nalidixic acid resistance to the host and resistance to T5 growth in such a host. The inhibition by nalidixic acid was also not due to a cell poison formed between nalidixic acid and the A subunit (K. N. Kreuzer and N. R. Cozzarelli, J. Bacteriol. 140:424-435, 1979) because nalidixic acid inhibited growth of T5 in a gyrA(Ts) mutant (KNK453) at 42 degrees C. We suggest that T5 grows in KNK453 at 42 degrees C because its gyrA(Ts) mutation is leaky for T5. Inhibition of T5 growth due to inactivation of host DNA gyrase was caused mainly by inhibition of T5 DNA replication. In addition, however, late T5 genes were barely expressed when host DNA gyrase was inactivated.

摘要

噬菌体T5在携带gyrB温度敏感突变[gyrB(Ts)]的大肠杆菌中于42℃的非允许温度下不能生长,但它在42℃时能在gyrA(Ts)突变体中生长。这些发现表明,宿主DNA促旋酶的A亚基不是必需的,而B亚基是T5生长所必需的。新生霉素和香豆霉素A1对T5生长有强烈抑制作用,从而证实了B亚基的必要性,它们特异性地干扰宿主DNA促旋酶B亚基的功能。然而,萘啶酸也强烈抑制T5生长,它特异性地干扰A亚基的功能。这种抑制是由于萘啶酸与A亚基的相互作用,而不仅仅是由于它与DNA的结合,因为宿主gyrA基因中的适当突变赋予宿主对萘啶酸的抗性以及在这样的宿主中对T5生长的抗性。萘啶酸的抑制作用也不是由于萘啶酸与A亚基之间形成的细胞毒素(K.N.克鲁泽和N.R.科扎雷利,《细菌学杂志》140:424 - 435,1979),因为萘啶酸在42℃时抑制了T5在gyrA(Ts)突变体(KNK453)中的生长。我们认为T5在42℃时能在KNK453中生长是因为其gyrA(Ts)突变对T5是渗漏的。宿主DNA促旋酶失活导致的T5生长抑制主要是由于T5 DNA复制受到抑制。然而,此外,当宿主DNA促旋酶失活时,T5晚期基因几乎不表达。

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