The crystal structure of the CRISPR-associated protein Csn2 from Streptococcus agalactiae

被引：18

作者：

Ellinger, Philipp ^{[2
]}

Arslan, Zihni ^{[3
]}

Wurm, Reinhild ^{[3
]}

Tschapek, Britta ^{[2
]}

MacKenzie, Colin ^{[4
,5
]}

Pfeffer, Klaus ^{[4
,5
]}

Panjikar, Santosh ^{[6
]}

Wagner, Rolf ^{[3
]}

Schmitt, Lutz ^{[2
]}

Gohlke, Holger ^{[1
]}

Pul, Uemit ^{[3
]}

Smits, Sander H. J. ^{[2
]}

机构：

[1] Univ Dusseldorf, Inst Pharmaceut & Med Chem, D-40225 Dusseldorf, Germany

[2] Univ Dusseldorf, Inst Biochem, D-40225 Dusseldorf, Germany

[3] Univ Dusseldorf, Inst Mol Biol Bacteria, D-40225 Dusseldorf, Germany

[4] Univ Dusseldorf, Inst Med Microbiol, D-40225 Dusseldorf, Germany

[5] Univ Dusseldorf, Hosp Hyg, D-40225 Dusseldorf, Germany

[6] DESY, EMBL Hamburg Outstn, D-22603 Hamburg, Germany

来源：

JOURNAL OF STRUCTURAL BIOLOGY | 2012年 / 178卷 / 03期

关键词：

CRISPR; Spacer integration; X-ray crystallography; MD-simulation; DNA binding; ANTIVIRAL DEFENSE; IMMUNE-SYSTEM; SMALL RNA; DOMAIN; BACTERIA; IDENTIFICATION; CONSERVATION; VALIDATION; SIMULATION; SEQUENCES;

D O I：

10.1016/j.jsb.2012.04.006

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 prokaryotic immune system, CRISPR, confers an adaptive and inheritable defense mechanism against invasion by mobile genetic elements. Guided by small CRISPR RNAs (crRNAs), a diverse family of CRISPR-associated (Cas) proteins mediates the targeting and inactivation of foreign DNA. Here, we demonstrate that Csn2, a Cas protein likely involved in spacer integration, forms a tetramer in solution and structurally possesses a ring-like structure. Furthermore, co-purified Ca2+ was found important for the DNA binding property of Csn2, which contains a helicase fold, with highly conserved DxD and RR motifs found throughout Csn2 proteins. We could verify that Csn2 binds ds-DNA. In addition molecular dynamics simulations suggested a Csn2 conformation that can "sit" on the DNA helix and binds DNA in a groove on the outside of the ring. (c) 2012 Elsevier Inc. All rights reserved.

引用

页码：350 / 362

页数：13

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