Type III CRISPR complexes from Thermus thermophilus

• Authors: Marta Szychowska , Wojciech Siwek , Damian Pawolski , Asgar Kazrani , Krzysztof Pyrc , Matthias Bochtler
• Languages of publication: EN

Abstracts

EN

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.

Keywords

EN

CRISPR-Cas; endonuclease; Csm; Cmr; Thermus thermophilus; crRNA; mass spectrometry (MS); chromatography

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2016
• Volume: 63
• Issue: 2
• Pages: 377-386

Dates

published

2016

received

2016-02-05

revised

2016-05-24

accepted

2016-05-24

(unknown)

2016-06-14

Contributors

author

Marta Szychowska

• International Institute of Molecular and Cell Biology, Warsaw, Poland
• Institute of Technical Biochemistry, Faculty of Biotechnology and Food Sciences, Technical University of Lodz, Łódź, Poland

author

Wojciech Siwek

• International Institute of Molecular and Cell Biology, Warsaw, Poland

author

Damian Pawolski

• International Institute of Molecular and Cell Biology, Warsaw, Poland

author

Asgar Kazrani

• International Institute of Molecular and Cell Biology, Warsaw, Poland

author

Krzysztof Pyrc

• Microbiology Department, Faculty of Biochemistry Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland
• Malopolska Centre of Biotechnology, Jagiellonian University, Kraków, Poland

author

Matthias Bochtler

• International Institute of Molecular and Cell Biology, Warsaw, Poland
• Institute of Biochemistry and Biophysics PAS, Warsaw, Poland

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Identifiers

DOI

10.18388/abp.2016_1261