Department of Veterinary and Animal Sciences, University of Copenhagen, Stigbøjlen 4, 1870 Frederiksberg C, Denmark.
Department of Biotechnology, Ghent University, Valentin Vaerwijckweg 1, 9000 Gent, Belgium.
Viruses. 2020 Oct 31;12(11):1247. doi: 10.3390/v12111247.
phages are divided into two genera; and , showing only limited intergenus homology. Here, we aim to identify the lytic genes of both genera using two representative phages (F352 and F379) from our collection. We performed a detailed in silico analysis searching for conserved protein domains and found that the predicted lytic genes are not organized into lysis cassettes but are conserved within each genus. To verify the function of selected lytic genes, the proteins were expressed in , followed by lytic assays. Our results show that phages encode a typical signal peptide (SP) endolysin dependent on the Sec-pathway for translocation and a holin for activation. In contrast, phages encode a novel endolysin that does not belong to currently described endolysin groups. This endolysin also uses the Sec-pathway for translocation but induces lysis of after overexpression. Interestingly, co-expression of this endolysin with an overlapping gene delayed and limited cell lysis, suggesting that this gene functions as a lysis inhibitor. These results indicate that phages regulate lysis timing by a yet undescribed mechanism. In conclusion, we found that the two phage genera control lysis by two distinct mechanisms.
噬菌体分为两个属; 和 ,它们之间只有有限的属间同源性。在这里,我们旨在使用来自我们收集的两个代表性噬菌体(F352 和 F379)来识别这两个属的裂解基因。我们进行了详细的计算机分析,搜索保守的蛋白结构域,发现预测的裂解基因没有组织成裂解盒,但在每个属内都保守。为了验证所选裂解基因的功能,将蛋白质在 中表达,然后进行裂解测定。我们的结果表明, 噬菌体编码一种典型的信号肽(SP)内溶素,该内溶素依赖于 Sec 途径进行易位,并编码一种参与激活的 holin。相比之下, 噬菌体编码一种新型内溶素,不属于目前描述的内溶素组。这种内溶素也使用 Sec 途径进行易位,但在过表达后诱导 的裂解。有趣的是,这种内溶素与重叠基因的共表达延迟并限制了细胞裂解,表明该基因作为裂解抑制剂发挥作用。这些结果表明, 噬菌体通过一种尚未描述的机制来调节裂解时间。总之,我们发现两种 噬菌体属通过两种不同的机制来控制裂解。