Structure and mechanism of the CMR complex for CRISPR-mediated antiviral immunity.

Structure and mechanism of the CMR complex for CRISPR-mediated antiviral immunity.
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DOI:
10.1016/j.molcel.2011.12.013
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发表时间:
2012-02-10
期刊:
影响因子:
16
通讯作者:
White, Malcolm F.
White, Malcolm F.
中科院分区:
生物学1区
文献类型:
--
作者:
Zhang, Jing;Rouillon, Christophe;Kerou, Melina;Reeks, Judith;Brugger, Kim;Graham, Shirley;Reimann, Julia;Cannone, Giuseppe;Liu, Huanting;Albers, Sonja-Verena;Naismith, James H.;Spagnolo, Laura;White, Malcolm F.

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原核常规间隔回文重复序列(CRISPR)系统利用基因组编码的CRISPR RNA (crRNA),来源于入侵病毒,并与CRISPR相关(CAS)蛋白结合到核糖核蛋白复合物中,在随后的感染中靶向和降解病毒DNA或RNA。RNA是CMR复合物的靶标。在solfataricus中,该复合物由7个CAS蛋白亚基(Cmr1-7)组成,并携带多种靶向crRNA的“有效载荷”。给出了Cmr7的晶体结构和配合物的低分辨率结构。S. solfataricus CMR在UA二核苷酸的核内分解反应中切割RNA靶标。这种活性依赖于crRNA中8个核苷酸重复衍生的5 '序列,而不依赖于靶标中原间隔相关基序(proto-spacer associated motif, PAM)的存在。靶标RNA和引导RNA都可以被切割,尽管单个引导RNA分子可以支持多个靶标的降解。
The prokaryotic Clusters of Regularly Interspaced Palindromic Repeats (CRISPR) system utilizes genomically-encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subsequent infection. RNA is targeted by the CMR complex. In Sulfolobus solfataricus, this complex is composed of seven CAS protein subunits (Cmr1-7) and carries a diverse “payload” of targeting crRNA. The crystal structure of Cmr7 and low resolution structure of the complex are presented. S. solfataricus CMR cleaves RNA targets in an endonucleolytic reaction at UA dinucleotides. This activity is dependent on the 8-nucleotide repeat-derived 5′ sequence in the crRNA, but not on the presence of a proto-spacer associated motif (PAM) in the target. Both target and guide RNAs can be cleaved, although a single molecule of guide RNA can support the degradation of multiple targets.
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