title: Escherichia coli DinB inhibits replication fork progression without significantly inducing the SOS response.
abstract: The SOS response is readily triggered by replication fork stalling caused by DNA damage or a dysfunctional replicative apparatus in Escherichia coli cells. E. coli dinB encodes DinB DNA polymerase and its expression is upregulated during the SOS response. DinB catalyzes translesion DNA synthesis in place of a replicative DNA polymerase III that is stalled at a DNA lesion. We showed previously that DNA replication was suppressed without exogenous DNA damage in cells overproducing DinB. In this report, we confirm that this was due to a dose-dependent inhibition of ongoing replication forks by DinB. Interestingly, the DinB-overproducing cells did not significantly induce the SOS response even though DNA replication was perturbed. RecA protein is activated by forming a nucleoprotein filament with single-stranded DNA, which leads to the onset of the SOS response. In the DinB-overproducing cells, RecA was not activated to induce the SOS response. However, the SOS response was observed after heat-inducible activation in strain recA441 (encoding a temperature-sensitive RecA) and after replication blockage in strain dnaE486 (encoding a temperature-sensitive catalytic subunit of the replicative DNA polymerase III) at a non-permissive temperature when DinB was overproduced in these cells. Furthermore, since catalytically inactive DinB could avoid the SOS response to a DinB-promoted fork block, it is unlikely that overproduced DinB takes control of primer extension and thus limits single-stranded DNA. These observations suggest that DinB possesses a feature that suppresses DNA replication but does not abolish the cell's capacity to induce the SOS response. We conclude that DinB impedes replication fork progression in a way that does not activate RecA, in contrast to obstructive DNA lesions and dysfunctional replication machinery.
authors: Division of Systems Biology, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan.
authors: Tetsuya
authors: T
authors: Mori
authors: Tatsuro
authors: T
authors: Nakamura
authors: Naoto
authors: N
authors: Okazaki
authors: Asako
authors: A
authors: Furukohri
authors: Hisaji
authors: H
authors: Maki
authors: Masahiro Tatsumi
authors: MT
authors: Akiyama
mesh_terms: D004249:DNA Damage; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism; D019951:DNA Polymerase beta / Q000235:genetics* / Q000378:metabolism; D004261:DNA Replication; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004277:DNA, Single-Stranded; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D046228:Microarray Analysis / Q000379:methods; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism; D013014:SOS Response, Genetics; D015854:Up-Regulation
chemical_list: D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; C094399:DinB protein, E coli; D029968:Escherichia coli Proteins; D011938:Rec A Recombinases; D004258:DNA Polymerase III; D019951:DNA Polymerase beta