5xvo

From Proteopedia

(Difference between revisions)

Current revision

E. fae Cas1-Cas2/prespacer/target ternary complex revealing DNA sampling and half-integration states

Structural highlights

5xvo is a 18 chain structure with sequence from Enterococcus faecalis TX0027. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 3.1Å
Ligands:
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Publication Abstract from PubMed

CRISPR (clustered regularly interspaced short palindromic repeats) and the nearby cas (CRISPR-associated) operon establish an RNA-based adaptive immunity system in prokaryotes1-5. Molecular memory is created when a short foreign DNA-derived prespacer is integrated into the CRISPR array as a new spacer6-9. Whereas the RNA-guided CRISPR interference mechanism varies widely among CRISPR-Cas systems, the spacer integration mechanism is essentially identical7-9. The conserved Cas1 and Cas2 proteins form an integrase complex consisting two distal Cas1 dimers bridged by a Cas2 dimer in the middle6,10. The prespacer is bound by Cas1-Cas2 as a dual forked DNA, and the terminal 3'-OH of each 3'-overhang serves as an attacking nucleophile during integration11-14. Importantly, the prespacer is preferentially integrated into the leader-proximal region of the CRISPR array1,7,10,15, guided by the leader sequence and a pair of inverted repeats (IRs) inside the CRISPR repeat7,15-20. Spacer integration in the most well-studied Escherichia coli Type I-E CRISPR system further relies on the bacterial Integration Host Factor (IHF)21,22. In Type II-A CRISPR, however, Cas1-Cas2 alone integrates spacer efficiently in vitro18; other Cas proteins (Cas9 and Csn2) play accessory roles in prespacer biogenesis17,23. Focusing on the Enterococcus faecalis Type II-A system24, here we report four structure snapshots of Cas1-Cas2 during spacer integration. EfaCas1-Cas2 selectively binds to a splayed 30-bp prespacer bearing 4-nt 3'-overhangs. Three molecular events take place upon encountering a target: Cas1-Cas2/prespacer first searches for half-sites stochastically, then preferentially interacts with the leader-side CRISPR repeat and catalyzes a nucleophilic attack that connects one strand of the leader-proximal repeat to the prespacer 3'-overhang. Recognition of the spacer half-site requires DNA bending and leads to full integration. We derive a mechanistic framework explaining the stepwise spacer integration process and the leader-proximal preference.

How type II CRISPR-Cas establish immunity through Cas1-Cas2-mediated spacer integration.,Xiao Y, Ng S, Nam KH, Ke A Nature. 2017 Sep 4. doi: 10.1038/nature24020. PMID:28869593^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Xiao Y, Ng S, Nam KH, Ke A. How type II CRISPR-Cas establish immunity through Cas1-Cas2-mediated spacer integration. Nature. 2017 Sep 4. doi: 10.1038/nature24020. PMID:28869593 doi:http://dx.doi.org/10.1038/nature24020

1 Structural highlights
2 Publication Abstract from PubMed
3 See Also
4 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/5xvo"

@@ Line 1: / Line 1: @@
 ==E. fae Cas1-Cas2/prespacer/target ternary complex revealing DNA sampling and half-integration states==
-<StructureSection load='5xvo' size='340' side='right' caption='[[5xvo]], [[Resolution|resolution]] 3.10&Aring;' scene=''>
+<StructureSection load='5xvo' size='340' side='right'caption='[[5xvo]], [[Resolution|resolution]] 3.10&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[5xvo]] is a 18 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5XVO OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5XVO FirstGlance]. <br>
+<table><tr><td colspan='2'>[[5xvo]] is a 18 chain structure with sequence from [https://en.wikipedia.org/wiki/Enterococcus_faecalis_TX0027 Enterococcus faecalis TX0027]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5XVO OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5XVO FirstGlance]. <br>
-</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></td></tr>
+</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 3.1&#8491;</td></tr>
-<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5xvn|5xvn]], [[5xvp|5xvp]]</td></tr>
+<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></td></tr>
-<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5xvo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5xvo OCA], [http://pdbe.org/5xvo PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5xvo RCSB], [http://www.ebi.ac.uk/pdbsum/5xvo PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5xvo ProSAT]</span></td></tr>
+<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5xvo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5xvo OCA], [https://pdbe.org/5xvo PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5xvo RCSB], [https://www.ebi.ac.uk/pdbsum/5xvo PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5xvo ProSAT]</span></td></tr>
 </table>
-== Function ==
-[[http://www.uniprot.org/uniprot/E6GPD7_ENTFL E6GPD7_ENTFL]] CRISPR (clustered regularly interspaced short palindromic repeat), is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). Acts as a dsDNA endonuclease. Involved in the integration of spacer DNA into the CRISPR cassette.[HAMAP-Rule:MF_01470] [[http://www.uniprot.org/uniprot/E6GPD6_ENTFL E6GPD6_ENTFL]] CRISPR (clustered regularly interspaced short palindromic repeat), is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain sequences complementary to antecedent mobile elements and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). Functions as a ssRNA-specific endoribonuclease. Involved in the integration of spacer DNA into the CRISPR cassette.[HAMAP-Rule:MF_01471]
 <div style="background-color:#fffaf0;">
 == Publication Abstract from PubMed ==
@@ Line 19: / Line 17: @@
 </div>
 <div class="pdbe-citations 5xvo" style="background-color:#fffaf0;"></div>
+==See Also==
+*[[Endonuclease 3D structures|Endonuclease 3D structures]]
 == References ==
 <references/>
 __TOC__
 </StructureSection>
-[[Category: Ke, A]]
+[[Category: Enterococcus faecalis TX0027]]
-[[Category: Nam, K H]]
+[[Category: Large Structures]]
-[[Category: Ng, S]]
+[[Category: Ke A]]
-[[Category: Xiao, Y]]
+[[Category: Nam KH]]
-[[Category: Ca]]
+[[Category: Ng S]]
-[[Category: Crispr]]
+[[Category: Xiao Y]]
-[[Category: Immune system]]

5xvo

From Proteopedia

Current revision

E. fae Cas1-Cas2/prespacer/target ternary complex revealing DNA sampling and half-integration states

Structural highlights

Publication Abstract from PubMed

See Also

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

Views

Personal tools

Navigation

Search

Toolbox