Sequences spanning the leader-repeat junction mediate CRISPR adaptation to phage in Streptococcus thermophilus.
Sequences spanning the leader-repeat junction mediate CRISPR adaptation to phage in Streptococcus thermophilus.
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DOI：
10.1093/nar/gku1407
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发表时间：
2015-02-18
影响因子：
14.9
通讯作者：
Terns MP
中科院分区：
文献类型：
--
作者：
Wei Y;Chesne MT;Terns RM;Terns MP
CRISPR-Cas系统是基于RNA的免疫系统，其保护原核生物免受入侵者如质粒和质粒的侵害。在适应中，即免疫应答的初始阶段，短的外源DNA片段被捕获并整合到宿主CRISPR基因座中，以提供针对遇到的外源核酸的遗传防御。每个CRISPR含有一个100 - 500 bp的前导元件，其通常包括转录启动子，随后是夹在相同的100 - 35 bp直接重复序列的拷贝之间的捕获的100 - 35 bp序列（间隔子）的阵列。新的分隔符将直接添加到引线的下游。在这里，我们分析了嗜热链球菌CRISPR1基因座对噬菌体感染的适应，以确定该过程所必需的顺式作用元件。我们表明，CRISPR基因座的前导序列和单个重复序列足以适应该系统。此外，我们确定了一个前导序列元素能够刺激适应在一个休眠重复。我们发现，在CRISPR的前导序列和重复序列中，整合位点10 bp内的序列是该过程所必需的。我们的研究结果表明，CRISPR前导序列-重复序列连接处的信息对于这种II-A型系统和可能的其他CRISPR-Cas系统的适应至关重要。
CRISPR-Cas systems are RNA-based immune systems that protect prokaryotes from invaders such as phages and plasmids. In adaptation, the initial phase of the immune response, short foreign DNA fragments are captured and integrated into host CRISPR loci to provide heritable defense against encountered foreign nucleic acids. Each CRISPR contains a ∼100–500 bp leader element that typically includes a transcription promoter, followed by an array of captured ∼35 bp sequences (spacers) sandwiched between copies of an identical ∼35 bp direct repeat sequence. New spacers are added immediately downstream of the leader. Here, we have analyzed adaptation to phage infection in Streptococcus thermophilus at the CRISPR1 locus to identify cis-acting elements essential for the process. We show that the leader and a single repeat of the CRISPR locus are sufficient for adaptation in this system. Moreover, we identified a leader sequence element capable of stimulating adaptation at a dormant repeat. We found that sequences within 10 bp of the site of integration, in both the leader and repeat of the CRISPR, are required for the process. Our results indicate that information at the CRISPR leader-repeat junction is critical for adaptation in this Type II-A system and likely other CRISPR-Cas systems.
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影响因子：
16
作者：
Hale, Caryn R.;Majumdar, Sonali;Elmore, Joshua;Pfister, Neil;Compton, Mark;Olson, Sara;Resch, Alissa M.;Glover, Claiborne V. C., III;Graveley, Brenton R.;Terns, Rebecca M.;Terns, Michael P.
通讯作者：
Terns, Michael P.
影响因子：
5.2
作者：
Bernick DL;Cox CL;Dennis PP;Lowe TM
通讯作者：
Lowe TM
影响因子：
3.2
作者：
Deveau, Helene;Barrangou, Rodolphe;Moineau, Sylvain
通讯作者：
Moineau, Sylvain
影响因子：
4.8
作者：
Ki Hyun Nam;Kurinov, Igor;Ke, Ailong
通讯作者：
Ke, Ailong
DOI：
10.1126/science.1192272
发表时间：
2010-09-10
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Haurwitz RE;Jinek M;Wiedenheft B;Zhou K;Doudna JA
通讯作者：
Doudna JA