Type III CRISPR complexes from Thermus thermophilus. | Zendy

Marta Szychowska | Zendy; Wojciech Siwek | Zendy; Damian Pawolski | Zendy; Asgar Abbas Kazrani | Zendy; Krzysztof Pyrć | Zendy; Matthias Bochtler | Zendy

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Type III CRISPR complexes from Thermus thermophilus.

Author(s) -

Marta Szychowska,

Wojciech Siwek,

Damian Pawolski,

Asgar Abbas Kazrani,

Krzysztof Pyrć,

Matthias Bochtler

Publication year - 2016

Publication title -

acta biochimica polonica

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.452

H-Index - 78

eISSN - 1734-154X

pISSN - 0001-527X

DOI - 10.18388/abp.2016_1261

Subject(s) - thermus thermophilus , trans activating crrna , crispr , biology , computational biology , chemistry , streptococcus thermophilus , genetics , palindrome , gene , bacteria , escherichia coli , lactobacillus

Pathogen-specific acquired immunity in bacteria is mediated by the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas systems. Thermus thermophilus strain HB8 contains CRISPR systems of several major subtypes (type I, IIIA and IIIB), and has become a widely studied model for CRISPR biology. We have selected two highly expressed CRISPR spacers, crRNA 2.1 and crRNA 2.2, and have enriched endogenous T. thermophilus proteins that co-purify with these crRNAs. Mass spectroscopy indicates that the chromatography protocol enriches predominantly Csm complex subunits, but also Cmr subunits. After several chromatographic steps, size exclusion chromatography indicated a molecular mass of the crRNA associated complex of 265±69 kDa. In agreement with earlier work, crRNAs of different lengths (containing the selected spacers) were observed. Most of these were completely lost when several T. thermophilus csm genes were ablated.

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