Kiljunen Saija, Hakala Kristo, Pinta Elise, Huttunen Suvi, Pluta Patrycja, Gador Aneta, Lönnberg Harri, Skurnik Mikael
Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki and Helsinki University Central Hospital Laboratory, Helsinki, Finland.
Department of Medical Biochemistry and Molecular Biology, Institute of Biomedicine, University of Turku, Turku, Finland.
Microbiology (Reading). 2005 Dec;151(Pt 12):4093-4102. doi: 10.1099/mic.0.28265-0.
Bacteriophage piR1-37 was isolated based on its ability to infect strain YeO3-R1, a virulence-plasmid-cured O antigen-negative derivative of Yersinia enterocolitica serotype O : 3. In this study, the phage receptor was found to be a structure in the outer core hexasaccharide of Y. enterocolitica O : 3 LPS. The phage receptor was present in the outer core of strains of many other Y. enterocolitica serotypes, but also in some Yersinia intermedia strains. Surprisingly, the receptor structure resided in the O antigen of Yersinia pseudotuberculosis O : 9. Electron microscopy demonstrated that phiR1-37 particles have an icosahedral head of 88 nm, a short neck of 10 nm, a long contractile tail of 236 nm, and tail fibres of at least 86 nm. This implies that the phage belongs to the order Caudovirales and the family Myoviridae in the ICTV (International Committee for Taxonomy of Viruses) classification. phiR1-37 was found to have a lytic life cycle, with eclipse and latent periods of 40 and 50 min, respectively, and a burst size of approximately 80 p.f.u. per infected cell. Restriction digestions and PFGE showed that the phiR1-37 genome was dsDNA and approximately 270 kb in size. Enzymically hydrolysed DNA was subjected to HPLC-MS/MS analysis, which demonstrated that the phiR1-37 genome is composed of DNA in which thymidine (T) is >99 % replaced by deoxyuridine (dU). The only organisms known to have similar DNA are the Bacillus subtilis-specific bacteriophages PBS1 and PBS2. N-terminal amino acid sequences of four major structural proteins did not show any similarity to (viral) protein sequences in databases, indicating that close relatives of phiR1-37 have not yet been characterized. Genes for two of the structural proteins, p24 and p46, were identified from the partially sequenced phiR1-37 genome.
噬菌体piR1-37是基于其感染YeO3-R1菌株的能力而分离得到的,YeO3-R1是小肠结肠炎耶尔森氏菌血清型O:3的一种无毒力质粒、O抗原阴性的衍生物。在本研究中,发现噬菌体受体是小肠结肠炎耶尔森氏菌O:3脂多糖外核心六糖中的一种结构。该噬菌体受体存在于许多其他小肠结肠炎耶尔森氏菌血清型菌株的外核心中,也存在于一些中间耶尔森氏菌菌株中。令人惊讶的是,受体结构存在于假结核耶尔森氏菌O:9的O抗原中。电子显微镜显示,phiR1-37颗粒有一个88nm的二十面体头部、一个10nm的短颈部、一个236nm的长收缩尾部和至少86nm的尾丝。这表明该噬菌体在国际病毒分类委员会(ICTV)的分类中属于有尾噬菌体目和肌尾噬菌体科。发现phiR1-37具有裂解性生命周期,潜伏期和隐蔽期分别为40分钟和50分钟,每个感染细胞的爆发量约为80个噬菌斑形成单位。限制性酶切和脉冲场凝胶电泳显示,phiR1-37基因组是双链DNA,大小约为270kb。对酶解后的DNA进行HPLC-MS/MS分析,结果表明phiR1-37基因组由胸腺嘧啶(T)>99%被脱氧尿苷(dU)取代的DNA组成。已知具有类似DNA的唯一生物是枯草芽孢杆菌特异性噬菌体PBS1和PBS2。四种主要结构蛋白的N端氨基酸序列与数据库中的(病毒)蛋白序列没有任何相似性,这表明phiR1-37的近亲尚未得到鉴定。从部分测序的phiR1-37基因组中鉴定出了两种结构蛋白p24和p46的基因。