Castilla-Llorente Virginia, Meijer Wilfried J J, Salas Margarita
Instituto de Biología Molecular 'Eladio Viñuela' (CSIC), Centro de Biología Molecular 'Severo Ochoa' (CSIC-UAM), Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain.
Nucleic Acids Res. 2009 Aug;37(15):4955-64. doi: 10.1093/nar/gkp504. Epub 2009 Jun 15.
Members of groups 1 (e.g. 29) and 2 (e.g. Nf) of the 29 family of phages infect the spore forming bacterium Bacillus subtilis. Although classified as lytic phages, the lytic cycle of 29 can be suppressed and its genome can become entrapped into the B. subtilis spore. This constitutes an alternative infection strategy that depends on the presence of binding sites for the host-encoded protein Spo0A in the 29 genome. Binding of Spo0A to these sites represses 29 transcription and prevents initiation of DNA replication. Although the Nf genome can also become trapped into B. subtilis spores, in vivo studies showed that its lytic cycle is less susceptible to spo0A-mediated suppression than that of 29. Here we have analysed the molecular mechanism underlying this difference showing that Spo0A differently affects transcription and replication initiation of the genomes of these phages. Thus, whereas Spo0A represses all three main early promoters of 29, it only represses one out of the three equivalent early promoters of Nf. In addition, contrary to 29, Spo0A does not prevent the in vitro initiation of Nf DNA replication. Altogether, the differences in Spo0A-mediated regulation of transcription and replication between 29 and Nf explain their different behaviours in vivo.
29 噬菌体家族的第 1 组(如 29 号)和第 2 组(如 Nf)成员感染形成芽孢的细菌枯草芽孢杆菌。尽管 29 噬菌体被归类为裂解性噬菌体,但其裂解周期可被抑制,其基因组可被困入枯草芽孢杆菌的芽孢中。这构成了一种替代感染策略,该策略取决于 29 噬菌体基因组中宿主编码蛋白 Spo0A 的结合位点的存在。Spo0A 与这些位点的结合会抑制 29 噬菌体的转录并阻止 DNA 复制的起始。尽管 Nf 噬菌体的基因组也可被困入枯草芽孢杆菌的芽孢中,但体内研究表明,其裂解周期比 29 噬菌体更不易受到 spo0A 介导的抑制。在这里,我们分析了这种差异背后的分子机制,结果表明 Spo0A 对这些噬菌体基因组的转录和复制起始的影响不同。因此,虽然 Spo0A 抑制 29 噬菌体的所有三个主要早期启动子,但它只抑制 Nf 噬菌体三个等效早期启动子中的一个。此外,与 29 噬菌体不同,Spo0A 并不阻止 Nf 噬菌体 DNA 在体外的复制起始。总之,29 噬菌体和 Nf 噬菌体在 Spo0A 介导的转录和复制调控方面的差异解释了它们在体内的不同行为。