Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation | |
Huo, YW; Nam, KH; Ding, F; Lee, HJ; Wu, LJ; Xiao, YB; Farchione, MD; Zhou, S; Rajashankar, K; Kurinov, I | |
刊名 | NATURE STRUCTURAL & MOLECULAR BIOLOGY
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2014 | |
卷号 | 21期号:9页码:771-777 |
通讯作者 | Ke, AL (reprint author), Cornell Univ, Dept Mol Biol & Genet, Ithaca, NY 14850 USA.,ak425@cornell.edu |
英文摘要 | CRISPR drives prokaryotic adaptation to invasive nucleic acids such as phages and plasmids, using an RNA-mediated interference mechanism. Interference in type I CRISPR-Cas systems requires a targeting Cascade complex and a degradation machine, Cas3, which contains both nuclease and helicase activities. Here we report the crystal structures of Thermobifida fusca Cas3 bound to single-stranded (ss) DNA substrate and show that it is an obligate 3'-to-5' ssDNase that preferentially accepts substrate directly from the helicase moiety. Conserved residues in the HD-type nuclease coordinate two irons for ssDNA cleavage: We demonstrate ATP coordination and conformational flexibility of the SF2-type helicase domain. Cas3 is specifically guided toward Cascade-bound target DNA by a PAM sequence, through physical interactions with both the nontarget substrate strand and the CasA protein. The sequence of recognition events ensures well-controlled DNA targeting and degradation of foreign DNA by Cascade and Cas3. |
学科主题 | Biochemistry & Molecular Biology; Biophysics; Cell Biology |
类目[WOS] | Biochemistry & Molecular Biology ; Biophysics ; Cell Biology |
关键词[WOS] | IN-VITRO RECONSTITUTION ; IMMUNE-SYSTEM ; INTERFERENCE ; HELICASE ; PROKARYOTES ; NUCLEASE ; REVEALS ; COMPLEX ; TARGET ; ENZYME |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000341404200006 |
内容类型 | 期刊论文 |
版本 | 出版稿 |
源URL | [http://202.127.25.143/handle/331003/225] ![]() |
专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
推荐引用方式 GB/T 7714 | Huo, YW,Nam, KH,Ding, F,et al. Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation[J]. NATURE STRUCTURAL & MOLECULAR BIOLOGY,2014,21(9):771-777. |
APA | Huo, YW.,Nam, KH.,Ding, F.,Lee, HJ.,Wu, LJ.,...&Ke, AL.(2014).Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation.NATURE STRUCTURAL & MOLECULAR BIOLOGY,21(9),771-777. |
MLA | Huo, YW,et al."Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation".NATURE STRUCTURAL & MOLECULAR BIOLOGY 21.9(2014):771-777. |
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