Département de Microbiologie, Unité Biologie Moléculaire du Gène chez les Extrêmophiles, Institut Pasteur, Paris, France.
Open Biol. 2013 Apr 17;3(4):130010. doi: 10.1098/rsob.130010.
The 2 465 177 bp genome of Sulfolobus islandicus LAL14/1, host of the model rudivirus SIRV2, was sequenced. Exhaustive comparative genomic analysis of S. islandicus LAL14/1 and the nine other completely sequenced S. islandicus strains isolated from Iceland, Russia and USA revealed a highly syntenic common core genome of approximately 2 Mb and a long hyperplastic region containing most of the strain-specific genes. In LAL14/1, the latter region is enriched in insertion sequences, CRISPR (clustered regularly interspaced short palindromic repeats), glycosyl transferase genes, toxin-antitoxin genes and MITE (miniature inverted-repeat transposable elements). The tRNA genes of LAL14/1 are preferential targets for the integration of mobile elements but clusters of atypical genes (CAG) are also integrated elsewhere in the genome. LAL14/1 carries five CRISPR loci with 10 per cent of spacers matching perfectly or imperfectly the genomes of archaeal viruses and plasmids found in the Icelandic hot springs. Strikingly, the CRISPR_2 region of LAL14/1 carries an unusually long 1.9 kb spacer interspersed between two repeat regions and displays a high similarity to pING1-like conjugative plasmids. Finally, we have developed a genetic system for S. islandicus LAL14/1 and created ΔpyrEF and ΔCRISPR_1 mutants using double cross-over and pop-in/pop-out approaches, respectively. Thus, LAL14/1 is a promising model to study virus-host interactions and the CRISPR/Cas defence mechanism in Archaea.
嗜酸热硫化叶菌 LAL14/1 的基因组大小为 2465177bp,其宿主是模式 rudivirus SIRV2。我们对嗜酸热硫化叶菌 LAL14/1 及其从冰岛、俄罗斯和美国分离得到的其他 9 株全序列已测序的菌株进行了全面的比较基因组分析,结果显示它们具有高度保守的约 2Mb 的共有核心基因组,以及一个富含大多数菌株特异性基因的超大增生区域。在 LAL14/1 中,后者区域富含插入序列、CRISPR(成簇的、规律间隔的短回文重复序列)、糖基转移酶基因、毒素-抗毒素基因和 MITE(微型反转录转座元件)。LAL14/1 的 tRNA 基因是移动元件整合的首选靶点,但也有基因簇(CAG)整合到基因组的其他部位。LAL14/1 有 5 个 CRISPR 基因座,其中 10%的间隔序列与在冰岛温泉中发现的古菌病毒和质粒的基因组完全或不完全匹配。引人注目的是,LAL14/1 的 CRISPR_2 区域携带一个异常长的 1.9kb 间隔序列,位于两个重复区之间,与 pING1 样可转移质粒具有高度相似性。最后,我们开发了嗜酸热硫化叶菌 LAL14/1 的遗传系统,并分别使用双交叉和 pop-in/pop-out 方法创建了 ΔpyrEF 和 ΔCRISPR_1 突变体。因此,LAL14/1 是研究病毒-宿主相互作用和古菌 CRISPR/Cas 防御机制的有前途的模型。