Naslov
Roles of PriA protein and double-strand DNA break repair functions in UV-induced restriction alleviation in Escherichia coli

Autori
Ivančić-Baće, Ivana ; Vlašić, Ignacija ; Čogelja-Čajo, Gordana ; Brčić-Kostić, Krunoslav ; Salaj-Šmic, Erika

Izvornik
Genetics (0016-6731) 174 (2006), 4; 2137-2149

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

Ključne riječi
lambda fag; EcoKI enzim; inhibicija restrikcije; PriA protein; RecBCD enzim
(lambda phage; EcoKI enzyme; restriction alleviation; PriA protein; RecBCD enzyme)

Sažetak
It has been widely considered that DNA modification protects the chromosome of bacteria E. coli K-12 against their own restriction-modification systems. Chromosomal DNA is protected from degradation by methylation of target sequences. However, when unmethylated target sequences are generated in the host chromosome, the endonuclease activity of the EcoKI restriction-modification enzyme is inactivated by the ClpXP protease and DNA is protected. This process is known as restriction alleviation (RA) and it can be induced by UV irradiation (UV-induced RA). It has been proposed that chromosomal unmethylated target sequences, a signal for the cell to protect its own DNA, can be generated by homologous recombination during the repair of damaged DNA. In this study, we wanted to further investigate the genetic requirements for recombination proteins involved in the generation of unmethylated target sequences. For this purpose, we monitored the alleviation of EcoKI restriction by measuring the survival of unmodified λ in UV-irradiated cells. Our genetic analysis showed that UV-induced RA is dependent on the excision repair protein UvrA, the RecA loading activity of the RecBCD enzyme, the primosome assembly activity of the PriA helicase and partially dependent on RecFOR proteins. Based on our results, we propose that unmethylated target sequences are generated at the D-loop by the strand exchange of two hemimethylated duplex DNA and subsequent initiation of DNA replication.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA