+検索条件
-Structure paper
タイトル | Structure reveals why genome folding is necessary for site-specific integration of foreign DNA into CRISPR arrays. |
---|---|
ジャーナル・号・ページ | Nat Struct Mol Biol, Vol. 30, Issue 11, Page 1675-1685, Year 2023 |
掲載日 | 2023年9月14日 |
著者 | Andrew Santiago-Frangos / William S Henriques / Tanner Wiegand / Colin C Gauvin / Murat Buyukyoruk / Ava B Graham / Royce A Wilkinson / Lenny Triem / Kasahun Neselu / Edward T Eng / Gabriel C Lander / Blake Wiedenheft / |
PubMed 要旨 | Bacteria and archaea acquire resistance to viruses and plasmids by integrating fragments of foreign DNA into the first repeat of a CRISPR array. However, the mechanism of site-specific integration ...Bacteria and archaea acquire resistance to viruses and plasmids by integrating fragments of foreign DNA into the first repeat of a CRISPR array. However, the mechanism of site-specific integration remains poorly understood. Here, we determine a 560-kDa integration complex structure that explains how Pseudomonas aeruginosa Cas (Cas1-Cas2/3) and non-Cas proteins (for example, integration host factor) fold 150 base pairs of host DNA into a U-shaped bend and a loop that protrude from Cas1-2/3 at right angles. The U-shaped bend traps foreign DNA on one face of the Cas1-2/3 integrase, while the loop places the first CRISPR repeat in the Cas1 active site. Both Cas3 proteins rotate 100 degrees to expose DNA-binding sites on either side of the Cas2 homodimer, which each bind an inverted repeat motif in the leader. Leader sequence motifs direct Cas1-2/3-mediated integration to diverse repeat sequences that have a 5'-GT. Collectively, this work reveals new DNA-binding surfaces on Cas2 that are critical for DNA folding and site-specific delivery of foreign DNA. |
リンク | Nat Struct Mol Biol / PubMed:37710013 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.48 Å |
構造データ | EMDB-29280, PDB-8flj: |
由来 |
|
キーワード | RECOMBINATION / DNA BINDING PROTEIN / HYDROLASE/DNA / DNA integration / maintenance of CRISPR repeat elements / IHF-DNA complex / Enzymes altering nucleic acid conformation / Site specific endodeoxyribonucleases: cleavage is not sequence specific / Nucleotidyltransferases / GO:0008301 / HYDROLASE-DNA complex |