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| | ==Crystal structure of the Cas1-Cas2 complex from Escherichia coli== | | ==Crystal structure of the Cas1-Cas2 complex from Escherichia coli== |
| - | <StructureSection load='4p6i' size='340' side='right' caption='[[4p6i]], [[Resolution|resolution]] 2.30Å' scene=''> | + | <StructureSection load='4p6i' size='340' side='right'caption='[[4p6i]], [[Resolution|resolution]] 2.30Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4p6i]] is a 6 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4P6I OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4P6I FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4p6i]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12]. The January 2015 RCSB PDB [https://pdb.rcsb.org/pdb/static.do?p=education_discussion/molecule_of_the_month/index.html Molecule of the Month] feature on ''Cascade and CRISPR'' by David Goodsell is [https://dx.doi.org/10.2210/rcsb_pdb/mom_2015_1 10.2210/rcsb_pdb/mom_2015_1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4P6I OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4P6I FirstGlance]. <br> |
| - | </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=4p6i FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4p6i OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4p6i RCSB], [http://www.ebi.ac.uk/pdbsum/4p6i PDBsum]</span></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]] 2.3Å</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=4p6i FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4p6i OCA], [https://pdbe.org/4p6i PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4p6i RCSB], [https://www.ebi.ac.uk/pdbsum/4p6i PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4p6i ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/CAS2_ECOLI CAS2_ECOLI]] 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 (By similarity).<ref>PMID:21255106</ref> [[http://www.uniprot.org/uniprot/CAS1_ECOLI CAS1_ECOLI]] 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). Cas1 is thought to be involved in CRISPR adaptation, the first stage of CRISPR immunity. Might be involved in the addition/removal of CRISPR spacers. Endonucleolytically cleaves linear ssRNA, ssDNA and short (34 base) dsDNA as well as branched DNA substrates such as Holliday junctions, replication forks and 5'-flap DNA substrates. Genetic interactions suggest Cas1 interacts with components of the RecBC and RuvB DNA repair systems.<ref>PMID:21255106</ref> | + | [https://www.uniprot.org/uniprot/CAS2_ECOLI CAS2_ECOLI] 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 (By similarity).<ref>PMID:21255106</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| | + | <div class="pdbe-citations 4p6i" style="background-color:#fffaf0;"></div> |
| | | | |
| | ==See Also== | | ==See Also== |
| - | *[[Endonuclease|Endonuclease]] | + | *[[Endonuclease 3D structures|Endonuclease 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Doudna, J A]] | + | [[Category: Cascade and CRISPR]] |
| - | [[Category: Kranzusch, P J]] | + | [[Category: Escherichia coli K-12]] |
| - | [[Category: Noeske, J]] | + | [[Category: Large Structures]] |
| - | [[Category: Nunez, J K]] | + | [[Category: RCSB PDB Molecule of the Month]] |
| - | [[Category: Crispr-associated protein]] | + | [[Category: Doudna JA]] |
| - | [[Category: Hydrolase]] | + | [[Category: Kranzusch PJ]] |
| - | [[Category: Nuclease]] | + | [[Category: Noeske J]] |
| | + | [[Category: Nunez JK]] |
| Structural highlights
Function
CAS2_ECOLI 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 (By similarity).[1]
Publication Abstract from PubMed
The initial stage of CRISPR-Cas immunity involves the integration of foreign DNA spacer segments into the host genomic CRISPR locus. The nucleases Cas1 and Cas2 are the only proteins conserved among all CRISPR-Cas systems, yet the molecular functions of these proteins during immunity are unknown. Here we show that Cas1 and Cas2 from Escherichia coli form a stable complex that is essential for spacer acquisition and determine the 2.3-A-resolution crystal structure of the Cas1-Cas2 complex. Mutations that perturb Cas1-Cas2 complex formation disrupt CRISPR DNA recognition and spacer acquisition in vivo. Active site mutants of Cas2, unlike those of Cas1, can still acquire new spacers, thus indicating a nonenzymatic role of Cas2 during immunity. These results reveal the universal roles of Cas1 and Cas2 and suggest a mechanism by which Cas1-Cas2 complexes specify sites of CRISPR spacer integration.
Cas1-Cas2 complex formation mediates spacer acquisition during CRISPR-Cas adaptive immunity.,Nunez JK, Kranzusch PJ, Noeske J, Wright AV, Davies CW, Doudna JA Nat Struct Mol Biol. 2014 May 4. doi: 10.1038/nsmb.2820. PMID:24793649[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Perez-Rodriguez R, Haitjema C, Huang Q, Nam KH, Bernardis S, Ke A, DeLisa MP. Envelope stress is a trigger of CRISPR RNA-mediated DNA silencing in Escherichia coli. Mol Microbiol. 2011 Feb;79(3):584-99. doi: 10.1111/j.1365-2958.2010.07482.x. Epub, 2010 Dec 13. PMID:21255106 doi:10.1111/j.1365-2958.2010.07482.x
- ↑ Nunez JK, Kranzusch PJ, Noeske J, Wright AV, Davies CW, Doudna JA. Cas1-Cas2 complex formation mediates spacer acquisition during CRISPR-Cas adaptive immunity. Nat Struct Mol Biol. 2014 May 4. doi: 10.1038/nsmb.2820. PMID:24793649 doi:http://dx.doi.org/10.1038/nsmb.2820
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