Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
Host nucleases generate prespacers for primed adaptation in the E. coli type I-E CRISPR-Cas system. / Shiriaeva, Anna A.; Kuznedelov, Konstanin; Fedorov, Ivan; Musharova, Olga; Khvostikov, Timofey; Tsoy, Yuliya; Kurilovich, Elena; Smith, Gerald R.; Semenova, Ekaterina; Severinov, Konstantin.
в: Science advances, Том 8, № 47, eabn8650, 11.2022.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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TY - JOUR
T1 - Host nucleases generate prespacers for primed adaptation in the E. coli type I-E CRISPR-Cas system
AU - Shiriaeva, Anna A.
AU - Kuznedelov, Konstanin
AU - Fedorov, Ivan
AU - Musharova, Olga
AU - Khvostikov, Timofey
AU - Tsoy, Yuliya
AU - Kurilovich, Elena
AU - Smith, Gerald R.
AU - Semenova, Ekaterina
AU - Severinov, Konstantin
N1 - Publisher Copyright: Copyright © 2022 The Authors, some rights reserved;
PY - 2022/11
Y1 - 2022/11
N2 - CRISPR-Cas systems provide prokaryotes with adaptive immunity against foreign nucleic acids. In Escherichia coli, immunity is acquired upon integration of 33-bp spacers into CRISPR arrays. DNA targets complementary to spacers get degraded and serve as a source of new spacers during a process called primed adaptation. Precursors of such spacers, prespacers, are ~33-bp double-stranded DNA fragments with a ~4-nt 3′ overhang. The mechanism of prespacer generation is not clear. Here, we use FragSeq and biochemical approaches to determine enzymes involved in generation of defined prespacer ends. We demonstrate that RecJ is the main exonuclease trimming 5′ ends of prespacer precursors, although its activity can be partially substituted by ExoVII. The RecBCD complex allows single strand–specific RecJ to process double-stranded regions flanking prespacers. Our results reveal intricate functional interactions of genome maintenance proteins with CRISPR interference and adaptation machineries during generation of prespacers capable of integration into CRISPR arrays.
AB - CRISPR-Cas systems provide prokaryotes with adaptive immunity against foreign nucleic acids. In Escherichia coli, immunity is acquired upon integration of 33-bp spacers into CRISPR arrays. DNA targets complementary to spacers get degraded and serve as a source of new spacers during a process called primed adaptation. Precursors of such spacers, prespacers, are ~33-bp double-stranded DNA fragments with a ~4-nt 3′ overhang. The mechanism of prespacer generation is not clear. Here, we use FragSeq and biochemical approaches to determine enzymes involved in generation of defined prespacer ends. We demonstrate that RecJ is the main exonuclease trimming 5′ ends of prespacer precursors, although its activity can be partially substituted by ExoVII. The RecBCD complex allows single strand–specific RecJ to process double-stranded regions flanking prespacers. Our results reveal intricate functional interactions of genome maintenance proteins with CRISPR interference and adaptation machineries during generation of prespacers capable of integration into CRISPR arrays.
UR - http://www.scopus.com/inward/record.url?scp=85142940341&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/4e62b218-c4dd-35e2-95bb-cf534ffa9136/
U2 - 10.1126/sciadv.abn8650
DO - 10.1126/sciadv.abn8650
M3 - Article
VL - 8
JO - Science advances
JF - Science advances
SN - 2375-2548
IS - 47
M1 - eabn8650
ER -
ID: 100595491