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| | ==Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex with bound target dsDNA== | | ==Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex with bound target dsDNA== |
| - | <StructureSection load='6b44' size='340' side='right' caption='[[6b44]], [[Resolution|resolution]] 2.90Å' scene=''> | + | <SX load='6b44' size='340' side='right' viewer='molstar' caption='[[6b44]], [[Resolution|resolution]] 2.90Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6b44]] is a 12 chain structure with sequence from [http://en.wikipedia.org/wiki/ ] and [http://en.wikipedia.org/wiki/Pseab Pseab]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6B44 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6B44 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6b44]] is a 12 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_aeruginosa Pseudomonas aeruginosa], [https://en.wikipedia.org/wiki/Pseudomonas_aeruginosa_UCBPP-PA14 Pseudomonas aeruginosa UCBPP-PA14] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6B44 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6B44 FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[6b45|6b45]], [[6b46|6b46]], [[6b47|6b47]], [[6b48|6b48]]</td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 2.9Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">csy1, PA14_33330 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=208963 PSEAB]), csy2, PA14_33320 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=208963 PSEAB]), csy3, csy1-3, PA14_33310 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=208963 PSEAB]), cas6f, csy4, PA14_33300 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=208963 PSEAB])</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=6b44 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6b44 OCA], [https://pdbe.org/6b44 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6b44 RCSB], [https://www.ebi.ac.uk/pdbsum/6b44 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6b44 ProSAT]</span></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=6b44 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6b44 OCA], [http://pdbe.org/6b44 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6b44 RCSB], [http://www.ebi.ac.uk/pdbsum/6b44 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6b44 ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://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> [[http://www.uniprot.org/uniprot/CSY1_PSEAB CSY1_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). Cas3 and Cascade participate in CRISPR interference, the third stage of CRISPR immunity (Potential). Involved in crRNA production or stability. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.<ref>PMID:21398535</ref> <ref>PMID:22522703</ref> [[http://www.uniprot.org/uniprot/CSY3_PSEAB CSY3_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). Cas3 and Cascade participate in CRISPR interference, the third stage of CRISPR immunity (Potential). Involved in crRNA production or stability. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.<ref>PMID:21398535</ref> <ref>PMID:22522703</ref> [[http://www.uniprot.org/uniprot/CSY2_PSEAB CSY2_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). Cas3 and Cascade participate in CRISPR interference, the third stage of CRISPR immunity (Potential). Absolutely required for crRNA production or stability. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.<ref>PMID:21398535</ref> <ref>PMID:22522703</ref> | + | [https://www.uniprot.org/uniprot/CSY1_PSEAB CSY1_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). Cas3 and Cascade participate in CRISPR interference, the third stage of CRISPR immunity (Potential). Involved in crRNA production or stability. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.<ref>PMID:21398535</ref> <ref>PMID:22522703</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </div> | | </div> |
| | <div class="pdbe-citations 6b44" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 6b44" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[CRISPR subtype I-F|CRISPR subtype I-F]] |
| | + | *[[Endonuclease 3D structures|Endonuclease 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| - | </StructureSection> | + | </SX> |
| - | [[Category: Pseab]] | + | [[Category: Large Structures]] |
| - | [[Category: Bartesaghi, A]] | + | [[Category: Pseudomonas aeruginosa]] |
| - | [[Category: Earl, L]] | + | [[Category: Pseudomonas aeruginosa UCBPP-PA14]] |
| - | [[Category: Falconieri, V]] | + | [[Category: Synthetic construct]] |
| - | [[Category: Fox, T]] | + | [[Category: Bartesaghi A]] |
| - | [[Category: Guo, T W]] | + | [[Category: Earl L]] |
| - | [[Category: Merk, A]] | + | [[Category: Falconieri V]] |
| - | [[Category: Patel, D J]] | + | [[Category: Fox T]] |
| - | [[Category: Rao, P]] | + | [[Category: Guo TW]] |
| - | [[Category: Subramaniam, S]] | + | [[Category: Merk A]] |
| - | [[Category: Yang, H]] | + | [[Category: Patel DJ]] |
| - | [[Category: Crispr-ca]]
| + | [[Category: Rao P]] |
| - | [[Category: Immune system-rna-dna complex]]
| + | [[Category: Subramaniam S]] |
| | + | [[Category: Yang H]] |
| Structural highlights
Function
CSY1_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). Cas3 and Cascade participate in CRISPR interference, the third stage of CRISPR immunity (Potential). Involved in crRNA production or stability. The Csy ribonucleoprotein complex binds target ssDNA with high affinity but target dsDNA with much lower affinity.[1] [2]
Publication Abstract from PubMed
Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. A central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or RNA for degradation in a sequence- and site-specific manner analogous to RNA interference. Although the complexes display considerable diversity in their composition and architecture, many basic mechanisms underlying target recognition and cleavage are highly conserved. Using cryoelectron microscopy (cryo-EM), we show that the binding of target double-stranded DNA (dsDNA) to a type I-F CRISPR system yersinia (Csy) surveillance complex leads to large quaternary and tertiary structural changes in the complex that are likely necessary in the pathway leading to target dsDNA degradation by a trans-acting helicase-nuclease. Comparison of the structure of the surveillance complex before and after dsDNA binding, or in complex with three virally encoded anti-CRISPR suppressors that inhibit dsDNA binding, reveals mechanistic details underlying target recognition and inhibition.
Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex.,Guo TW, Bartesaghi A, Yang H, Falconieri V, Rao P, Merk A, Eng ET, Raczkowski AM, Fox T, Earl LA, Patel DJ, Subramaniam S Cell. 2017 Oct 5;171(2):414-426.e12. doi: 10.1016/j.cell.2017.09.006. PMID:28985564[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ 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
- ↑ Guo TW, Bartesaghi A, Yang H, Falconieri V, Rao P, Merk A, Eng ET, Raczkowski AM, Fox T, Earl LA, Patel DJ, Subramaniam S. Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex. Cell. 2017 Oct 5;171(2):414-426.e12. doi: 10.1016/j.cell.2017.09.006. PMID:28985564 doi:http://dx.doi.org/10.1016/j.cell.2017.09.006
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