Cas1-Cas2 complex formation mediates spacer acquisition during CRISPR-Cas adaptive immunity

被引：350

作者：

Nunez, James K. ^{[1
]}

Kranzusch, Philip J. ^{[1
,2
]}

Noeske, Jonas ^{[1
]}

Wright, Addison V. ^{[1
]}

Davies, Christopher W. ^{[1
]}

Doudna, Jennifer A. ^{[1
,2
,3
,4
]}

机构：

[1] Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA

[2] Univ Calif Berkeley, Howard Hughes Med Inst, Berkeley, CA 94720 USA

[3] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA

[4] Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Phys Biosci Div, Berkeley, CA 94720 USA

来源：

NATURE STRUCTURAL & MOLECULAR BIOLOGY | 2014年 / 21卷 / 06期

基金：

美国国家科学基金会;

关键词：

ANTIVIRUS IMMUNITY; STRUCTURAL BASIS; RNA; DEFENSE; PROTEIN; SYSTEM; ENDORIBONUCLEASE; ENDONUCLEASE; SEQUENCES;

D O I：

10.1038/nsmb.2820

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The initial stage of CRISPR-Cas immunity involves the integration of foreign DNA spacer segments into the host genomic CRISPR locus. The nucleases Cas1 and Cas2 are the only proteins conserved among all CRISPR-Cas systems, yet the molecular functions of these proteins during immunity are unknown. Here we show that Cas1 and Cas2 from Escherichia coli form a stable complex that is essential for spacer acquisition and determine the 2.3-angstrom-resolution crystal structure of the Cas1-Cas2 complex. Mutations that perturb Cas1-Cas2 complex formation disrupt CRISPR DNA recognition and spacer acquisition in vivo. Active site mutants of Cas2, unlike those of Cas1, can still acquire new spacers, thus indicating a nonenzymatic role of Cas2 during immunity. These results reveal the universal roles of Cas1 and Cas2 and suggest a mechanism by which Cas1-Cas2 complexes specify sites of CRISPR spacer integration.

引用

页码：528 / 534

页数：7

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