Uchiyama Jumpei, Takemura-Uchiyama Iyo, Kato Shin-Ichiro, Takeuchi Hiroaki, Sakaguchi Yoshihiko, Ujihara Takako, Daibata Masanori, Shimakura Hidekatsu, Okamoto Noriaki, Sakaguchi Masahiro, Matsuzaki Shigenobu
Department of Veterinary Microbiology, School of Veterinary Medicine, Azabu University, Kanagawa 252-5201, Japan.
Research Institute of Molecular Genetics, Kochi University, Kochi 783-8502, Japan.
FEMS Microbiol Lett. 2016 Aug;363(16). doi: 10.1093/femsle/fnw157. Epub 2016 Jul 7.
We have recently reported the active Helicobacter pylori bacteriophages (phages), KHP30 and KHP40, the genomic DNAs of which exist as episomes in host bacterial strains isolated in Japan (i.e. pseudolysogeny). In this study, we examined the possibility of the lysogeny of active KHP30-like phages in Japanese H. pylori strains, because their genomes contain a putative integrase gene. Only the NY40 strain yielded partial detection of a KHP30-like prophage sequence in PCR among 174 Japanese H. pylori isolates, except for strains producing the above active phages. Next, according to the genomic analysis of the NY40 strain, the KHP30-like prophage sequence was found to be located from ca. 524 to 549 kb in the host chromosome. The attachment sites, attL and attR, in the NY40 genome showed almost the same genomic location and sequence as those detected in a French isolate B38, suggesting that an active parental KHP30-like phage had integrated into the ancestral NY40 genome in a site-specific manner. The prophage found in the NY40 genome was assumed to have been genetically modified, after site-specific integration. These, together with the data in the KHP30-like prophages of other H. pylori genomes, suggest that the lysogenic state of the KHP30-like phages is generally unstable.
我们最近报道了活性幽门螺杆菌噬菌体KHP30和KHP40,其基因组DNA在日本分离的宿主细菌菌株中以附加体形式存在(即假溶原性)。在本研究中,我们研究了活性KHP30样噬菌体在日本幽门螺杆菌菌株中发生溶原性的可能性,因为它们的基因组包含一个假定的整合酶基因。在174株日本幽门螺杆菌分离株中,除了产生上述活性噬菌体的菌株外,只有NY40菌株在PCR中部分检测到KHP30样前噬菌体序列。接下来,根据NY40菌株的基因组分析,发现KHP30样前噬菌体序列位于宿主染色体中约524至549 kb处。NY40基因组中的附着位点attL和attR显示出与法国分离株B38中检测到的几乎相同的基因组位置和序列,这表明活性亲本KHP30样噬菌体已以位点特异性方式整合到祖先NY40基因组中。在NY40基因组中发现的前噬菌体被认为在位点特异性整合后发生了基因修饰。这些结果,连同其他幽门螺杆菌基因组中KHP30样前噬菌体的数据,表明KHP30样噬菌体的溶原状态通常是不稳定的。