Abstract

The uvrB gene belongs to the SOS network, encoding a key component of the nucleotide excision repair. The uvrB promoter region contains three identified promoters with four LexA binding sites, one consensus and six potential DnaA binding sites. A more than threefold increase in transcription of the chromosomal uvrB gene is observed in both the ΔlexA ΔsulA cells and dnaAA345S cells, and a fivefold increase in the ΔlexA ΔsulA dnaAA345S cells relative to the wild-type cells. The full activity of the uvrB promoter region requires both the uvrBp1-2 and uvrBp3 promoters and is repressed by both the DnaA and LexA proteins. LexA binds tightly to LexA-box1 at the uvrBp1-2 promoter irrespective of the presence of DnaA and this binding is important for the control of the uvrBp1-2 promoter. DnaA and LexA, however, compete for binding to and regulation of the uvrBp3 promoter in which the DnaA-box6 overlaps with LexA-box4. The transcription control of uvrBp3 largely depends on DnaA-box6. Transcription of other SOS regulon genes, such as recN and dinJ, is also repressed by both DnaA and LexA. Interestingly, the absence of LexA in the presence of the DnaAA345S mutant leads to production of elongated cells with incomplete replication, aberrant nucleoids and slow growth. We propose that DnaA is a modulator for maintenance of genome integrity during the SOS response by limiting the expression of the SOS regulon.

Highlights

  • The uvrB gene encodes the UvrB protein, one of the key components of the nucleotide excision repair (NER) system (Truglio et al, 2006)

  • The result suggests that DnaA could directly be involved in control of the uvrB gene expression since DnaAA345S binds to DnaA-boxes with a lower affinity compared to wild-type DnaA

  • The uvrB gene belongs to the SOS regulon which is regulated by LexA (Howard-Flanders et al, 1966)

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Summary

Introduction

The uvrB gene encodes the UvrB protein, one of the key components of the NER system (Truglio et al, 2006). SOS is a global response to DNA damage in which RecA filaments bound on ssDNA promote self-cleavage of the LexA protein. Cleavage of LexA induces expression of the SOS genes, resulting in an arrest of cell division for the time required to repair the damages (Walter, 1996). LexA regulates the Abbreviations: ADP/ATP-DnaA, ADP or ATP binds DnaA; bla, ampicillin resistance gene; cat, chloramphenicol resistance gene; DARS, DnaA reactivation site; DnaA-box, DnaA binding site; FRT, FLP recognition target; LexA-box, LexA binding site; neo, kanamycin resistance gene; NER, nucleotide excision repair; RIDA, regulatory inactivation of DnaA; ssDNA, single strand DNA; tet, tetracycline resistant gene

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