Structural basis for dimer formation of the CRISPR-associated protein Csm2 of Thermotoga maritima

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dc.citation.issue4
dc.citation.volume283
dc.contributor.authorGallo, Gloria [UNIFESP]
dc.contributor.authorAugusto, Gilles [UNIFESP]
dc.contributor.authorRangel, Giulliana [UNIFESP]
dc.contributor.authorZelanis, Andre [UNIFESP]
dc.contributor.authorMori, Marcelo A. [UNIFESP]
dc.contributor.authorCampos, Claudia B. [UNIFESP]
dc.contributor.authorWuertele, Martin [UNIFESP]
dc.coverageHoboken
dc.date.accessioned2020-11-03T14:40:28Z
dc.date.available2020-11-03T14:40:28Z
dc.date.issued2016
dc.description.abstractThe clusters of regularly interspaced short palindromic repeats (CRISPR) and the Cas (CRISPR-associated) proteins form an adaptive immune system in bacteria and archaea that evolved as an RNA-guided interference mechanism to target and degrade foreign genetic elements. In the so-called type IIIA CRISPR-Cas systems, Cas proteins from the Csm family form a complex of RNPs that are involved in surveillance and targeting tasks. In the present study, we report the crystal structure of Thermotoga maritima Csm2. This protein is considered to assemble into the helically shaped Csm RNP complex in a site opposite to the CRISPR RNA binding backbone. Csm2 was solved via cadmium single wavelength anomalous diffraction phasing at 2.4 angstrom resolution. The structure reveals that Csm2 is composed of a large 42 amino-acid long -helix flanked by three shorter -helices. The structure also shows that the protein is capable of forming dimers mainly via an extensive contact surface conferred by its long -helix. This interaction is further stabilized by the N-terminal helix, which is inserted into the C-terminal helical portion of the adjacent subunit. The dimerization of Csm2 was additionally confirmed by size exclusion chromatography of the pure recombinant protein followed by MS analysis of the eluted fractions. Because of its role in the assembly and functioning of the Csm CRISPR RNP complex, the crystal structure of Csm2 is of great importance for clarifying the mechanism of action of the subtype IIIA CRISPR-Cas system, as well as the similarities and diversities between the different CRISPR-Cas system. DatabaseThe structure of Thermotoga maritima Csm2 has been deposited in the Protein Data Bank under accession code .en
dc.description.affiliationUniv Fed Sao Paulo, Dept Sci & Technol, Rua Talim 330, BR-12231280 Sao Jose Dos Campos, Brazil
dc.description.affiliationLETA, Appl Toxinol Lab, Sao Paulo, Brazil
dc.description.affiliationInst Butantan, Ctr Toxins Immune Response & Cell Signaling CeTIC, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Dept Biophys, Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Sci & Technol, Rua Talim 330, BR-12231280 Sao Jose Dos Campos, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Biophys, Sao Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Sao Paulo Research Foundation)
dc.description.sponsorshipCNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico, National Council for Scientific and Technological Development, Brazil)
dc.description.sponsorshipIDFAPESP: 11/50963-4
dc.description.sponsorshipIDCNPq: 480411/2011-5
dc.description.sponsorshipIDCNPq: 448833/2014-0
dc.format.extent694-703
dc.identifierhttps://doi.org/10.1111/febs.13621
dc.identifier.citationFebs Journal. Hoboken, v. 283, n. 4, p. 694-703, 2016.
dc.identifier.doi10.1111/febs.13621
dc.identifier.issn1742-464X
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/58585
dc.identifier.wosWOS:000371071900010
dc.language.isoeng
dc.publisherWiley-Blackwell
dc.relation.ispartofFebs Journal
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectCd-SADen
dc.subjectCRISPR-Casen
dc.subjectcrRNPen
dc.subjectCsm2en
dc.subjectThermotoga maritimaen
dc.titleStructural basis for dimer formation of the CRISPR-associated protein Csm2 of Thermotoga maritimaen
dc.typeinfo:eu-repo/semantics/article
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