Department of Genetics and Molecular Biology, Faculty of Sciences, University of Pécs, H-7601 Pécs, Hungary.
J Bacteriol. 2010 Mar;192(6):1617-23. doi: 10.1128/JB.01335-09. Epub 2010 Jan 15.
Genes encoding the tail proteins of the temperate phage 16-3 of the symbiotic nitrogen-fixing bacterium Sinorhizobium meliloti 41 have been identified. First, a new host range gene, designated hII, was localized by using missense mutations. The corresponding protein was shown to be identical to the 85-kDa tail protein by determining its N-terminal sequence. Electron microscopic analysis showed that phage 16-3 possesses an icosahedral head and a long, noncontractile tail characteristic of the Siphoviridae. By using a lysogenic S. meliloti 41 strain, mutants with insertions in the putative tail region of the genome were constructed and virion morphology was examined after induction of the lytic cycle. Insertions in ORF017, ORF018a, ORF020, ORF021, the previously described h gene, and hII resulted in uninfectious head particles lacking tail structures, suggesting that the majority of the genes in this region are essential for tail formation. By using different bacterial mutants, it was also shown that not only the RkpM and RkpY proteins but also the RkpZ protein of the host takes part in the formation of the phage receptor. Results for the host range phage mutants and the receptor mutant bacteria suggest that the HII tail protein interacts with the capsular polysaccharide of the host and that the tail protein encoded by the original h gene recognizes a proteinaceous receptor.
已鉴定出与共生固氮菌根瘤菌(Sinorhizobium meliloti)41 温和噬菌体 16-3 的尾部蛋白编码基因。首先,通过使用错义突变,定位了一个新的宿主范围基因,称为 hII。通过确定其 N 端序列,证明相应的蛋白质与 85kDa 尾部蛋白相同。电子显微镜分析表明,噬菌体 16-3 具有二十面体头部和长而不可收缩的尾巴,这是 Siphoviridae 的特征。通过使用溶源的 S. meliloti 41 菌株,构建了基因组中假定尾部区域插入突变体,并在诱导裂解周期后检查了病毒形态。ORF017、ORF018a、ORF020、ORF021、先前描述的 h 基因和 hII 中的插入导致无传染性的头部颗粒缺少尾部结构,表明该区域的大多数基因对于尾部形成是必需的。通过使用不同的细菌突变体,还表明不仅宿主的 RkpM 和 RkpY 蛋白,而且 RkpZ 蛋白都参与噬菌体受体的形成。宿主范围噬菌体突变体和受体突变体细菌的结果表明,HII 尾部蛋白与宿主的荚膜多糖相互作用,并且原始 h 基因编码的尾部蛋白识别蛋白质受体。