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

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

Author Gallo, Gloria Autor UNIFESP Google Scholar
Augusto, Gilles Autor UNIFESP Google Scholar
Rangel, Giulliana Autor UNIFESP Google Scholar
Zelanis, Andre Autor UNIFESP Google Scholar
Mori, Marcelo A. Autor UNIFESP Google Scholar
Campos, Claudia B. Autor UNIFESP Google Scholar
Wuertele, Martin Autor UNIFESP Google Scholar
Abstract The 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 .
Keywords Cd-SAD
CRISPR-Cas
crRNP
Csm2
Thermotoga maritima
xmlui.dri2xhtml.METS-1.0.item-coverage Hoboken
Language English
Sponsor FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Sao Paulo Research Foundation)
CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico, National Council for Scientific and Technological Development, Brazil)
Grant number FAPESP: 11/50963-4
CNPq: 480411/2011-5
CNPq: 448833/2014-0
Date 2016
Published in Febs Journal. Hoboken, v. 283, n. 4, p. 694-703, 2016.
ISSN 1742-464X (Sherpa/Romeo, impact factor)
Publisher Wiley-Blackwell
Extent 694-703
Origin https://doi.org/10.1111/febs.13621
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000371071900010
URI https://repositorio.unifesp.br/handle/11600/58450

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