期刊
NUCLEIC ACIDS RESEARCH
卷 44, 期 22, 页码 10849-10861出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkw914
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资金
- National Institutes of Health (NIH) [GM10407, 5P20RR02437, GM097330]
- Russian Science Foundation [14-14-00988]
- Ministry of Education and Science of the Russian Federation [14.B25.31.0004]
- Russian Foundation for Basic Research [16-04-00767]
- Department of Energy [DE-SC0012518]
- M.J. Murdock Charitable Trust
- Charles and Johanna Busch Memorial Fund
- Natural Sciences and Engineering research Council of Canada (NSERC) [RGPIN-2015-04745]
- Skolkovo Institute of Science and Technology
- Charles and Johanna Busch predoctoral fellowship
- Russan Science Foundation [14-14-00988]
- U.S. Department of Energy (DOE) [DE-SC0012518] Funding Source: U.S. Department of Energy (DOE)
The Escherichia coli type I-E CRISPR-Cas system Cascade effector is a multisubunit complex that binds CRISPR RNA (crRNA). Through its 32-nucleotide spacer sequence, Cascade-bound crRNA recognizes protospacers in foreign DNA, causing its destruction during CRISPR interference or acquisition of additional spacers in CRISPR array during primed CRISPR adaptation. Within Cascade, the crRNA spacer interacts with a hexamer of Cas7 sub-units. We show that crRNAs with a spacer length reduced to 14 nucleotides cause primed adaptation, while crRNAs with spacer lengths of more than 20 nucleotides cause both primed adaptation and target interference in vivo. Shortened crRNAs assemble into altered-stoichiometry Cascade effector complexes containing less than the normal amount of Cas7 subunits. The results show that Cascade assembly is driven by crRNA and suggest that multisubunit type I CRISPR effectors may have evolved from much simpler ancestral complexes.
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