Buttimer Colin, Bottacini Francesca, Shkoporov Andrey N, Draper Lorraine A, Ross Paul, Hill Colin
APC Microbiome Ireland, University College Cork, T12 YT20 Cork, Ireland.
Department of Biological Sciences, Munster Technological University, T12 P928 Cork, Ireland.
Microorganisms. 2022 Jan 17;10(1):195. doi: 10.3390/microorganisms10010195.
is an anaerobic, high GC, Gram-positive bacillus commonly found in the human digestive tract that belongs to the class Coriobacteriia of the phylum Actinobacteria. This species has been of increasing interest as an important player in the metabolism of xenobiotics and dietary compounds. However, little is known regarding its susceptibility to bacteriophage predation and how this may influence its fitness. Here, we report the isolation of seven novel strains using cefotaxime and ceftriaxone as selective agents. We conducted comparative and pangenome analyses of these strains and those publicly available to investigate the diversity of prophages associated with this species. Prophage gene products represent a minimum of 5.8% of the pangenome, comprising at least ten distantly related prophage clades that display limited homology to currently known bacteriophages. All clades possess genes implicated in virion structure, lysis, lysogeny and, to a limited extent, DNA replication. Some prophages utilise tyrosine recombinases and diversity generating retroelements to generate phase variation among targeted genes. The prophages have differing levels of sensitivity to the CRISPR/cas systems of their hosts, with spacers from 44 isolates found to target only five out of the ten identified prophage clades. Furthermore, using a PCR-based approach targeting the prophage site, we were able to determine that several of these elements can excise from the host chromosome, thus supporting the notion that these are active prophages. The findings of this study provide further insights into the diversity of prophages infecting species of the phylum Actinobacteria.
是一种厌氧、高GC含量的革兰氏阳性杆菌,常见于人类消化道,属于放线菌门的放线杆菌纲。作为异生物质和膳食化合物代谢中的重要参与者,该物种越来越受到关注。然而,关于其对噬菌体捕食的敏感性以及这可能如何影响其适应性,人们知之甚少。在这里,我们报告了使用头孢噻肟和头孢曲松作为选择剂分离出七株新菌株。我们对这些菌株以及公开可用的菌株进行了比较和泛基因组分析,以研究与该物种相关的前噬菌体的多样性。前噬菌体基因产物占泛基因组的至少5.8%,包括至少十个与目前已知噬菌体同源性有限的远缘相关前噬菌体分支。所有分支都拥有与病毒体结构、裂解、溶原性以及在有限程度上与DNA复制相关的基因。一些前噬菌体利用酪氨酸重组酶和多样性产生逆转元件在靶向基因之间产生相变。前噬菌体对其宿主的CRISPR/cas系统具有不同程度的敏感性,从44个分离株中发现的间隔序列仅靶向十个已鉴定的前噬菌体分支中的五个。此外,使用基于PCR的方法靶向前噬菌体位点,我们能够确定其中几个元件可以从宿主染色体上切除,从而支持这些是活跃前噬菌体的观点。这项研究的结果为感染放线菌门物种的前噬菌体的多样性提供了进一步的见解。