CRISPR RNA-guided surveillance complex with Cas3 bound in its pre-nicking state
試料
複合体: CRISPR RNA-guided surveillance complex with Cas3 bound in its pre-nicking state
タンパク質・ペプチド: CRISPR-associated helicase, Cas3 family
タンパク質・ペプチド: CRISPR-associated protein, Cse1 family
タンパク質・ペプチド: CRISPR-associated protein, Cse4 family
タンパク質・ペプチド: Uncharacterized protein
RNA: crRNA
DNA: Target strand
タンパク質・ペプチド: CRISPR-associated protein, Cas5e family
DNA: Nontarget strand
タンパク質・ペプチド: CRISPR-associated protein, Cse3 family
リガンド: FE (III) ION
キーワード
CRISPR-Cas / Cascade / Cas3 / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA-RNA complex
機能・相同性
機能・相同性情報
nuclease activity / maintenance of CRISPR repeat elements / defense response to virus / RNA helicase activity / RNA binding / ATP binding / metal ion binding / identical protein binding 類似検索 - 分子機能
: / Helicase Cas3, CRISPR-associated, core / Cas3, HD domain / CRISPR-associated protein, CT1975 / CRISPR-associated protein Cse2 / Cse2 superfamily / CT1975-like protein / CRISPR-associated protein Cse2 (CRISPR_cse2) / CRISPR-associated protein, CasD / CRISPR-associated protein Cse1 ...: / Helicase Cas3, CRISPR-associated, core / Cas3, HD domain / CRISPR-associated protein, CT1975 / CRISPR-associated protein Cse2 / Cse2 superfamily / CT1975-like protein / CRISPR-associated protein Cse2 (CRISPR_cse2) / CRISPR-associated protein, CasD / CRISPR-associated protein Cse1 / CRISPR-associated protein Cse1 (CRISPR_cse1) / CRISPR-associated protein Cse3 / : / CRISPR associated protein / CRISPR_assoc / CRISPR-associated Cas3-type HD domain / CRISPR-associated Cas3-type HD domain superfamily / CRISPR-associated nuclease/helicase Cas3, C-terminal / HD Cas3-type domain profile. / CRISPR-associated protein, Cas5 / CRISPR-associated protein (Cas_Cas5) / CRISPR-associated protein Cas5, N-terminal / helicase superfamily c-terminal domain / Helicase, C-terminal / P-loop containing nucleoside triphosphate hydrolase 類似検索 - ドメイン・相同性
CRISPR-associated helicase, Cas3 family / CRISPR-associated protein, Cse1 family / CRISPR-associated protein, Cse2 family / CRISPR-associated protein, Cse4 family / CRISPR-associated protein, Cas5e family / CRISPR-associated protein, Cse3 family 類似検索 - 構成要素
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM118174
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM102543
米国
引用
ジャーナル: Science / 年: 2018 タイトル: Structure basis for RNA-guided DNA degradation by Cascade and Cas3. 著者: Yibei Xiao / Min Luo / Adam E Dolan / Maofu Liao / Ailong Ke / 要旨: Type I CRISPR-Cas system features a sequential target-searching and degradation process on double-stranded DNA by the RNA-guided Cascade (CRISPR associated complex for antiviral defense) complex and ...Type I CRISPR-Cas system features a sequential target-searching and degradation process on double-stranded DNA by the RNA-guided Cascade (CRISPR associated complex for antiviral defense) complex and the nuclease-helicase fusion enzyme Cas3, respectively. Here, we present a 3.7-angstrom-resolution cryo-electron microscopy (cryo-EM) structure of the Type I-E Cascade/R-loop/Cas3 complex, poised to initiate DNA degradation. Cas3 distinguishes Cascade conformations and only captures the R-loop-forming Cascade, to avoid cleaving partially complementary targets. Its nuclease domain recruits the nontarget strand (NTS) DNA at a bulged region for the nicking of single-stranded DNA. An additional 4.7-angstrom-resolution cryo-EM structure captures the postnicking state, in which the severed NTS retracts to the helicase entrance, to be threaded for adenosine 5'-triphosphate-dependent processive degradation. These snapshots form the basis for understanding RNA-guided DNA degradation in Type I-E CRISPR-Cas systems.