Journal Article

CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III

Vogel, Jörg, Sharma, Cynthia M., Charpentier, Emmanuelle, Eckert, Maria R., Chylinski, Krzysztof, Chao, Yanjie, Deltcheva, Elitza, Pirzada, Zaid A., Gonzales, Karine

2011

Overview

CRISPR/Cas systems constitute a widespread class of immunity systems that protect bacteria and archaea against phages and plasmids, and commonly use repeat/spacer-derived short crRNAs to silence foreign nucleic acids in a sequence-specific manner. Although the maturation of crRNAs represents a key event in CRISPR activation, the responsible endoribonucleases (CasE, Cas6, Csy4) are missing in many CRISPR/Cas subtypes. Here, differential RNA sequencing of the human pathogen Streptococcus pyogenes uncovered tracrRNA, a trans -encoded small RNA with 24-nucleotide complementarity to the repeat regions of crRNA precursor transcripts. We show that tracrRNA directs the maturation of crRNAs by the activities of the widely conserved endogenous RNase III and the CRISPR-associated Csn1 protein; all these components are essential to protect S. pyogenes against prophage-derived DNA. Our study reveals a novel pathway of small guide RNA maturation and the first example of a host factor (RNase III) required for bacterial RNA-mediated immunity against invaders. An alternative route to CRISPR-induced immunity CRISPR is a microbial RNA-based immune system protecting against viral and plasmid invasions. The CRISPR system is thought to rely on cleavage of a precursor RNA transcript by Cas endonucleases, but not all species with CRISPR-type immunity encode Cas proteins. A new study reveals an alternative pathway for CRISPR activation in the human pathogen Streptococcus pyogenes , in which a trans -encoded small RNA directs processing of precursor RNA into crRNAs through endogenous RNase III and the CRISPR-associated Csn1 protein. CRISPR is a microbial RNA-based immune system protecting against viral and plasmid invasions. The CRISPR system is thought to rely on cleavage of a precursor RNA transcript by Cas endonucleases, but not all species possessing CRISPR-type immunity encode Cas proteins. This study now describes an alternative pathway in Streptococcus pyogenes that employs trans -encoded small RNA that directs the processing of precursor RNA into crRNAs through endogenous RNase III and the CRISPR-associated Csn1 protein.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/326/41/1969 / 631/326/421 / 631/337/1645 / Archaea / Bacteria / Bacterial Proteins - chemistry / Bacterial Proteins - genetics / Bacterial Proteins - immunology / Bacterial Proteins - metabolism / Bacteriology / Biological and medical sciences / Conserved Sequence / DNA, Viral - genetics / DNA, Viral - metabolism / E coli / Escherichia coli / Fundamental and applied biological sciences. Psychology / Genetic aspects / Genetics / Humanities and Social Sciences / Microbiology / Models, Biological / multidisciplinary / Nucleic acids / Physiological aspects / Plasmids / Polymerase chain reaction / Prophages - genetics / Proteins / Ribonuclease / Ribonuclease III - metabolism / RNA / RNA Precursors - genetics / RNA Precursors - metabolism / RNA Processing, Post-Transcriptional / RNA, Bacterial - biosynthesis / RNA, Bacterial - genetics / RNA, Bacterial - immunology / RNA, Bacterial - metabolism / RNA, Small Untranslated / Science / Science (multidisciplinary) / Streptococcus pyogenes / Streptococcus pyogenes - genetics / Streptococcus pyogenes - immunology / Streptococcus pyogenes - metabolism / Streptococcus pyogenes - virology