Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation

	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
	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.