Ceyssens Pieter-Jan, Mesyanzhinov Vadim, Sykilinda Nina, Briers Yves, Roucourt Bart, Lavigne Rob, Robben Johan, Domashin Artem, Miroshnikov Konstantin, Volckaert Guido, Hertveldt Kirsten
Division of Gene Technology, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, Leuven B-3001, Belgium.
J Bacteriol. 2008 Feb;190(4):1429-35. doi: 10.1128/JB.01441-07. Epub 2007 Dec 7.
Pseudomonas aeruginosa phage YuA (Siphoviridae) was isolated from a pond near Moscow, Russia. It has an elongated head, encapsulating a circularly permuted genome of 58,663 bp, and a flexible, noncontractile tail, which is terminally and subterminally decorated with short fibers. The YuA genome is neither Mu- nor lambda-like and encodes 78 gene products that cluster in three major regions involved in (i) DNA metabolism and replication, (ii) host interaction, and (iii) phage particle formation and host lysis. At the protein level, YuA displays significant homology with phages M6, phiJL001, 73, B3, DMS3, and D3112. Eighteen YuA proteins were identified as part of the phage particle by mass spectrometry analysis. Five different bacterial promoters were experimentally identified using a promoter trap assay, three of which have a sigma54-specific binding site and regulate transcription in the genome region involved in phage particle formation and host lysis. The dependency of these promoters on the host sigma54 factor was confirmed by analysis of an rpoN mutant strain of P. aeruginosa PAO1. At the DNA level, YuA is 91% identical to the recently (July 2007) annotated phage M6 of the Lindberg typing set. Despite this level of DNA homology throughout the genome, both phages combined have 15 unique genes that do not occur in the other phage. The genome organization of both phages differs substantially from those of the other known Pseudomonas-infecting Siphoviridae, delineating them as a distinct genus within this family.
铜绿假单胞菌噬菌体YuA(长尾噬菌体科)从俄罗斯莫斯科附近的一个池塘中分离得到。它有一个细长的头部,包裹着一个58,663 bp的环状排列基因组,还有一条灵活的、不可收缩的尾部,尾部末端和亚末端装饰有短纤维。YuA基因组既不像Mu噬菌体也不像λ噬菌体,它编码78种基因产物,这些产物聚集在三个主要区域,分别参与(i)DNA代谢和复制,(ii)宿主相互作用,以及(iii)噬菌体颗粒形成和宿主裂解。在蛋白质水平上,YuA与噬菌体M6、phiJL001、73、B3、DMS3和D3112显示出显著的同源性。通过质谱分析鉴定出18种YuA蛋白是噬菌体颗粒的一部分。通过启动子捕获试验实验鉴定出5种不同的细菌启动子,其中3种具有sigma54特异性结合位点,并在参与噬菌体颗粒形成和宿主裂解的基因组区域调节转录。通过对铜绿假单胞菌PAO1的rpoN突变株的分析,证实了这些启动子对宿主sigma54因子的依赖性。在DNA水平上,YuA与最近(2007年7月)注释的林德伯格分型组噬菌体M6有91%的同一性。尽管整个基因组有这种程度的DNA同源性,但这两种噬菌体共有15个独特的基因,这些基因在另一种噬菌体中不存在。这两种噬菌体的基因组组织与其他已知感染假单胞菌的长尾噬菌体科的基因组组织有很大不同,将它们划定为该科内一个独特的属。