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High-throughput screen reveals sRNAs regulating crRNA biogenesis by targeting CRISPR leader to repress Rho termination

Author

Listed:
  • Ping Lin

    (Institute of Surgery Research, Daping Hospital, Army Medical University
    University of North Dakota)

  • Qinqin Pu

    (University of North Dakota)

  • Qun Wu

    (University of North Dakota
    Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine Shanghai)

  • Chuanmin Zhou

    (University of North Dakota)

  • Biao Wang

    (University of North Dakota)

  • Jacob Schettler

    (University of North Dakota)

  • Zhihan Wang

    (University of North Dakota)

  • Shugang Qin

    (University of North Dakota)

  • Pan Gao

    (University of North Dakota)

  • Rongpeng Li

    (Jiangsu Normal University)

  • Guoping Li

    (Southwestern Medical University, Pulmonary and Allergy Institute, Affiliated Hospital)

  • Zhenyu Cheng

    (Dalhousie University)

  • Lefu Lan

    (Chinese Academy of Sciences)

  • Jianxin Jiang

    (Institute of Surgery Research, Daping Hospital, Army Medical University)

  • Min Wu

    (University of North Dakota)

Abstract

Discovery of CRISPR-Cas systems is one of paramount importance in the field of microbiology. Currently, how CRISPR-Cas systems are finely regulated remains to be defined. Here we use small regulatory RNA (sRNA) library to screen sRNAs targeting type I-F CRISPR-Cas system through proximity ligation by T4 RNA ligase and find 34 sRNAs linking to CRISPR loci. Among 34 sRNAs for potential regulators of CRISPR, sRNA pant463 and PhrS enhance CRISPR loci transcription, while pant391 represses their transcription. We identify PhrS as a regulator of CRISPR-Cas by binding CRISPR leaders to suppress Rho-dependent transcription termination. PhrS-mediated anti-termination facilitates CRISPR locus transcription to generate CRISPR RNA (crRNA) and subsequently promotes CRISPR-Cas adaptive immunity against bacteriophage invasion. Furthermore, this also exists in type I-C/-E CRISPR-Cas, suggesting general regulatory mechanisms in bacteria kingdom. Our findings identify sRNAs as important regulators of CRISPR-Cas, extending roles of sRNAs in controlling bacterial physiology by promoting CRISPR-Cas adaptation priming.

Suggested Citation

  • Ping Lin & Qinqin Pu & Qun Wu & Chuanmin Zhou & Biao Wang & Jacob Schettler & Zhihan Wang & Shugang Qin & Pan Gao & Rongpeng Li & Guoping Li & Zhenyu Cheng & Lefu Lan & Jianxin Jiang & Min Wu, 2019. "High-throughput screen reveals sRNAs regulating crRNA biogenesis by targeting CRISPR leader to repress Rho termination," Nature Communications, Nature, vol. 10(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-11695-8
    DOI: 10.1038/s41467-019-11695-8
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    Cited by:

    1. Julian Trouillon & Kook Han & Ina Attrée & Stephen Lory, 2022. "The core and accessory Hfq interactomes across Pseudomonas aeruginosa lineages," Nature Communications, Nature, vol. 13(1), pages 1-16, December.

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