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| | ==Crystal structure of the Csy4-crRNA complex, orthorhombic form== | | ==Crystal structure of the Csy4-crRNA complex, orthorhombic form== |
| - | <StructureSection load='2xlk' size='340' side='right' caption='[[2xlk]], [[Resolution|resolution]] 1.81Å' scene=''> | + | <StructureSection load='2xlk' size='340' side='right'caption='[[2xlk]], [[Resolution|resolution]] 1.81Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[2xlk]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Pseab Pseab]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2XLK OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2XLK FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[2xlk]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_aeruginosa_UCBPP-PA14 Pseudomonas aeruginosa UCBPP-PA14]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2XLK OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2XLK FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2xli|2xli]], [[2xlj|2xlj]]</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]] 1.805Å</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=2xlk FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2xlk OCA], [http://pdbe.org/2xlk PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=2xlk RCSB], [http://www.ebi.ac.uk/pdbsum/2xlk PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=2xlk 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=2xlk FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2xlk OCA], [https://pdbe.org/2xlk PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2xlk RCSB], [https://www.ebi.ac.uk/pdbsum/2xlk PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2xlk ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/CAS6_PSEAB CAS6_PSEAB] 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). Processes pre-crRNA into individual crRNA units. Absolutely required for crRNA production or stability. Upon expression in E.coli endonucleolytically processes pre-crRNA, although disruption and reconstitution experiments indicate that in situ other genes are also required for processing. Yields 5'-hydroxy and 3'-phosphate groups. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.<ref>PMID:20829488</ref> <ref>PMID:21398535</ref> <ref>PMID:22522703</ref> |
| | == Evolutionary Conservation == | | == Evolutionary Conservation == |
| | [[Image:Consurf_key_small.gif|200px|right]] | | [[Image:Consurf_key_small.gif|200px|right]] |
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| | ==See Also== | | ==See Also== |
| | *[[CRISPR subtype I-F|CRISPR subtype I-F]] | | *[[CRISPR subtype I-F|CRISPR subtype I-F]] |
| - | *[[Endonuclease|Endonuclease]] | + | *[[Endonuclease 3D structures|Endonuclease 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Pseab]] | + | [[Category: Large Structures]] |
| - | [[Category: Doudna, J A]] | + | [[Category: Pseudomonas aeruginosa UCBPP-PA14]] |
| - | [[Category: Haurwitz, R E]] | + | [[Category: Doudna JA]] |
| - | [[Category: Jinek, M]] | + | [[Category: Haurwitz RE]] |
| - | [[Category: Wiedenheft, B]] | + | [[Category: Jinek M]] |
| - | [[Category: Zhou, K]] | + | [[Category: Wiedenheft B]] |
| - | [[Category: Crispr]]
| + | [[Category: Zhou K]] |
| - | [[Category: Endoribonuclease]]
| + | |
| - | [[Category: Hydrolase-rna complex]]
| + | |
| Structural highlights
Function
CAS6_PSEAB 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). Processes pre-crRNA into individual crRNA units. Absolutely required for crRNA production or stability. Upon expression in E.coli endonucleolytically processes pre-crRNA, although disruption and reconstitution experiments indicate that in situ other genes are also required for processing. Yields 5'-hydroxy and 3'-phosphate groups. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.[1] [2] [3]
Evolutionary Conservation
Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.
Publication Abstract from PubMed
Many bacteria and archaea contain clustered regularly interspaced short palindromic repeats (CRISPRs) that confer resistance to invasive genetic elements. Central to this immune system is the production of CRISPR-derived RNAs (crRNAs) after transcription of the CRISPR locus. Here, we identify the endoribonuclease (Csy4) responsible for CRISPR transcript (pre-crRNA) processing in Pseudomonas aeruginosa. A 1.8 angstrom crystal structure of Csy4 bound to its cognate RNA reveals that Csy4 makes sequence-specific interactions in the major groove of the crRNA repeat stem-loop. Together with electrostatic contacts to the phosphate backbone, these enable Csy4 to bind selectively and cleave pre-crRNAs using phylogenetically conserved serine and histidine residues in the active site. The RNA recognition mechanism identified here explains sequence- and structure-specific processing by a large family of CRISPR-specific endoribonucleases.
Sequence- and structure-specific RNA processing by a CRISPR endonuclease.,Haurwitz RE, Jinek M, Wiedenheft B, Zhou K, Doudna JA Science. 2010 Sep 10;329(5997):1355-8. PMID:20829488[4]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Haurwitz RE, Jinek M, Wiedenheft B, Zhou K, Doudna JA. Sequence- and structure-specific RNA processing by a CRISPR endonuclease. Science. 2010 Sep 10;329(5997):1355-8. PMID:20829488 doi:10.1126/science.1192272
- ↑ Cady KC, O'Toole GA. Non-identity-mediated CRISPR-bacteriophage interaction mediated via the Csy and Cas3 proteins. J Bacteriol. 2011 Jul;193(14):3433-45. doi: 10.1128/JB.01411-10. Epub 2011 Mar, 11. PMID:21398535 doi:http://dx.doi.org/10.1128/JB.01411-10
- ↑ Haurwitz RE, Sternberg SH, Doudna JA. Csy4 relies on an unusual catalytic dyad to position and cleave CRISPR RNA. EMBO J. 2012 Apr 20. doi: 10.1038/emboj.2012.107. PMID:22522703 doi:10.1038/emboj.2012.107
- ↑ Haurwitz RE, Jinek M, Wiedenheft B, Zhou K, Doudna JA. Sequence- and structure-specific RNA processing by a CRISPR endonuclease. Science. 2010 Sep 10;329(5997):1355-8. PMID:20829488 doi:10.1126/science.1192272
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