Sökning: onr:"swepub:oai:gup.ub.gu.se/315292" >
Structural and bioc...
Structural and biochemical characterization of in vivo assembled Lactococcus lactis CRISPR-Csm complex
-
- Sridhara, Sagar, 1989 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi,Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
-
Rai, J. (författare)
-
Whyms, C. (författare)
-
visa fler...
-
Goswami, H. (författare)
-
He, H. (författare)
-
Woodside, W. (författare)
-
Terns, M. P. (författare)
-
Li, H. (författare)
-
visa färre...
-
(creator_code:org_t)
- 2022-03-29
- 2022
- Engelska.
-
Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 5:1
- Relaterad länk:
-
https://gup.ub.gu.se...
-
visa fler...
-
https://doi.org/10.1...
-
visa färre...
Abstract
Ämnesord
Stäng
- The small RNA-mediated immunity in bacteria depends on foreign RNA-activated and self RNA-inhibited enzymatic activities. The multi-subunit Type III-A CRISPR-Cas effector complex (Csm) exemplifies this principle and is in addition regulated by cellular metabolites such as divalent metals and ATP. Recognition of the foreign or cognate target RNA (CTR) triggers its single-stranded deoxyribonuclease (DNase) and cyclic oligoadenylate (cOA) synthesis activities. The same activities remain dormant in the presence of the self or non-cognate target RNA (NTR) that differs from CTR only in its 3 '-protospacer flanking sequence (3 '-PFS). Here we employ electron cryomicroscopy (cryoEM), functional assays, and comparative cross-linking to study in vivo assembled mesophilic Lactococcus lactis Csm (LlCsm) at the three functional states: apo, the CTR- and the NTR-bound. Unlike previously studied Csm complexes, we observed binding of 3 '-PFS to Csm in absence of bound ATP and analyzed the structures of the four RNA cleavage sites. Interestingly, comparative crosslinking results indicate a tightening of the Csm3-Csm4 interface as a result of CTR but not NTR binding, reflecting a possible role of protein dynamics change during activation. This study presents Cryo-EM structures of the Lactococcus lactis Type III-A CRISPR-Cas effector complex in three functional states, including bound forms with cognate and non-cognate target RNA. The results suggest changing protein dynamics during effector complex activation.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Nyckelord
- crystal-structure
- gene-regulation
- rna cleavage
- cas system
- DNA
- classification
- microscopy
- evolution
- bacteria
- immunity
- Life Sciences & Biomedicine - Other Topics
- Science & Technology - Other
- Topics
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas