Naslov
Cas1–Cas2 physically and functionally interacts with DnaK to modulate CRISPR Adaptation

Autori
Killelea, Tom ; Dimude, Juachi U. ; He, Liu ; Stewart, Alison L. ; Kemm, Fiona E. ; Radovčić, Marin ; Ivančić-Baće, Ivana ; Rudolph, Christian J. ; Bolt, Edward L.

Izvornik
Nucleic acids research (0305-1048) 51 (2023); 1-13

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

Ključne riječi
adaptation, Cas1-Cas2, CRISPR-Cas, E. coli, DnaK

Sažetak
Prokaryotic Cas1–Cas2 protein complexes gener- ate adaptive immunity to mobile genetic elements (MGEs), by capture and integration of MGE DNA in to CRISPR sites. De novo immunity relies on naive adaptation––Cas1–Cas2 targeting of MGE DNA with- out the aid of pre-existing immunity ‘interference’ complexes––by mechanisms that are not clear. Us- ing E. coli we show that the chaperone DnaK inhibits DNA binding and integration by Cas1–Cas2, and in- hibits naive adaptation in cells that results from chro- mosomal self-targeting. Inhibition of naive adapta- tion was reversed by deleting DnaK from cells, by mutation of the DnaK substrate binding domain, and by expression of an MGE (phage ) protein. We also imaged fluorescently labelled Cas1 in living cells, ob- serving that Cas1 foci depend on active DNA replica- tion, and are much increased in frequency in cells lacking DnaK. We discuss a model in which DnaK provides a mechanism for restraining naive adapta- tion from DNA self-targeting, until DnaK is triggered to release Cas1–Cas2 to target MGE DNA.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA