Burmistrz Michal, Rodriguez Martinez Jose Ignacio, Krochmal Daniel, Staniec Dominika, Pyrc Krzysztof
Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.
Malopolska Center of Biotechnology, Jagiellonian University, Cracow, Poland.
J Bacteriol. 2017 Oct 31;199(23). doi: 10.1128/JB.00275-17. Print 2017 Dec 1.
The CRISPR-Cas (clustered regularly interspaced short palindromic repeat-CRISPR-associated protein) system is unique to prokaryotes and provides the majority of bacteria and archaea with immunity against nucleic acids of foreign origin. CRISPR RNAs (crRNAs) are the key element of this system, since they are responsible for its selectivity and effectiveness. Typical crRNAs consist of a spacer sequence flanked with 5' and 3' handles originating from repeat sequences that are important for recognition of these small RNAs by the Cas machinery. In this investigation, we studied the type I-C CRISPR-Cas system in , a human pathogen associated with periodontitis, rheumatoid arthritis, cardiovascular disease, and aspiration pneumonia. We demonstrated the importance of the 5' handle for crRNA recognition by the effector complex and consequently activity, as well as secondary trimming of the 3' handle, which was not affected by modifications of the repeat sequence., a clinically relevant Gram-negative, anaerobic bacterium, is one of the major etiologic agents of periodontitis and has been linked with the development of other clinical conditions, including rheumatoid arthritis, cardiovascular disease, and aspiration pneumonia. The presented results on the biogenesis and functions of crRNAs expand our understanding of CRISPR-Cas cellular defenses in and of horizontal gene transfer in bacteria.
CRISPR-Cas(成簇规律间隔短回文重复序列- CRISPR相关蛋白)系统是原核生物所特有的,为大多数细菌和古生菌提供针对外源核酸的免疫能力。CRISPR RNA(crRNA)是该系统的关键元件,因为它们决定了系统的选择性和有效性。典型的crRNA由一个间隔序列组成,其两侧是来自重复序列的5'和3'端序列,这些序列对于Cas机制识别这些小RNA很重要。在本研究中,我们研究了与牙周炎、类风湿性关节炎、心血管疾病和吸入性肺炎相关的人类病原体中的I-C型CRISPR-Cas系统。我们证明了5'端序列对于效应复合物识别crRNA进而发挥活性的重要性,以及3'端序列的二次修剪,这不受重复序列修饰的影响。是一种临床相关的革兰氏阴性厌氧菌,是牙周炎的主要病原体之一,并与包括类风湿性关节炎、心血管疾病和吸入性肺炎在内的其他临床病症的发展有关。所呈现的关于crRNA生物合成和功能的结果扩展了我们对中CRISPR-Cas细胞防御以及细菌中水平基因转移的理解。