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Title | Structure Studies of the CRISPR-Csm Complex Reveal Mechanism of Co-transcriptional Interference. |
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Journal, issue, pages | Cell, Vol. 176, Issue 1-2, Page 239-253.e16, Year 2019 |
Publish date | Jan 10, 2019 |
Authors | Lilan You / Jun Ma / Jiuyu Wang / Daria Artamonova / Min Wang / Liang Liu / Hua Xiang / Konstantin Severinov / Xinzheng Zhang / Yanli Wang / |
PubMed Abstract | Csm, a type III-A CRISPR-Cas interference complex, is a CRISPR RNA (crRNA)-guided RNase that also possesses target RNA-dependent DNase and cyclic oligoadenylate (cOA) synthetase activities. However, ...Csm, a type III-A CRISPR-Cas interference complex, is a CRISPR RNA (crRNA)-guided RNase that also possesses target RNA-dependent DNase and cyclic oligoadenylate (cOA) synthetase activities. However, the structural features allowing target RNA-binding-dependent activation of DNA cleavage and cOA generation remain unknown. Here, we report the structure of Csm in complex with crRNA together with structures of cognate or non-cognate target RNA bound Csm complexes. We show that depending on complementarity with the 5' tag of crRNA, the 3' anti-tag region of target RNA binds at two distinct sites of the Csm complex. Importantly, the interaction between the non-complementary anti-tag region of cognate target RNA and Csm1 induces a conformational change at the Csm1 subunit that allosterically activates DNA cleavage and cOA generation. Together, our structural studies provide crucial insights into the mechanistic processes required for crRNA-meditated sequence-specific RNA cleavage, RNA target-dependent non-specific DNA cleavage, and cOA generation. |
External links | Cell / PubMed:30503210 / PubMed Central |
Methods | EM (single particle) / X-ray diffraction |
Resolution | 2.9 - 3.8 Å |
Structure data | EMDB-9653, PDB-6ifk: EMDB-9654, PDB-6ifl: EMDB-9655: EMDB-9656, PDB-6ifr: EMDB-9657, PDB-6ifu: EMDB-9658, PDB-6ify: EMDB-9659, PDB-6ifz: EMDB-9660, PDB-6ig0: PDB-6ifn: |
Chemicals | ChemComp-ANP: ChemComp-MG: ChemComp-ZN: ChemComp-MN: ChemComp-ATP: |
Source |
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Keywords | RNA BINDING PROTEIN / Csm complex / Type III-A / CRISPR-Cas system |