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Pregled bibliografske jedinice broj: 967425

CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases.


(HRZZ, BBSRC) Radovčić, Marin; Killela, Tom; Savitskaya, Ekaterina; Wettstein, Lukas; Bolt, Edward L., Ivančić-Baće, Ivana
CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases. // Nucleic acids research, 46 (2018), 19; 10173-10183 doi:10.1093/nar/gky799 (međunarodna recenzija, članak, znanstveni)


Naslov
CRISPR-Cas adaptation in Escherichia coli requires RecBCD helicase but not nuclease activity, is independent of homologous recombination, and is antagonized by 5' ssDNA exonucleases.

Autori
Radovčić, Marin ; Killela, Tom ; Savitskaya, Ekaterina ; Wettstein, Lukas ; Bolt, Edward L., Ivančić-Baće, Ivana

Kolaboracija
HRZZ, BBSRC

Izvornik
Nucleic acids research (0305-1048) 46 (2018), 19; 10173-10183

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
CRISPR-Cas ; RecBCD ; Cas1-Cas2 ; nucleases ; E. coli

Sažetak
Prokaryotic adaptive immunity is established against mobile genetic elements (MGEs) by “naïve adaptation” when DNA fragments from a newly encountered MGE are integrated into CRISPR-Cas systems. In E. coli, DNA integration catalysed by Cas1-Cas2 integrase is well understood in mechanistic and structural detail but much less is known about events prior to integration that generate DNA for capture by Cas1-Cas2. Naïve adaptation in E. coli is thought to depend on the DNA helicase-nuclease RecBCD for generating DNA fragments for capture by Cas1-Cas2. The genetics presented here show that naïve adaptation does not require RecBCD nuclease activity but that helicase activity may be important. RecA loading by RecBCD inhibits adaptation explaining previously observed adaptation phenotypes that implicated RecBCD nuclease activity. Genetic analysis of other E. coli nucleases and naïve adaptation revealed that 5’ ssDNA tailed DNA molecules promote new spacer acquisition. We show that purified E. coli Cas1-Cas2 complex binds to and nicks 5’ ssDNA tailed duplexes and propose that E. coli Cas1-Cas2 nuclease activity on such DNA structures supports naïve adaptation.

Izvorni jezik
Engleski

Znanstvena područja
Biologija



POVEZANOST RADA


Projekt / tema
HRZZ-IP-2016-06-8861 - Cas3 kao kontrolna točka obrane CRISPR-Cas: razjašnjenje njegove regulacije istraživanjem stabilnosti proteina i prepisivanja u bakteriji Escherichia coli (Ivana Ivančić Baće, )

Ustanove
Prirodoslovno-matematički fakultet, Zagreb

Časopis indeksira:


  • Current Contents Connect (CCC)
  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE


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