Two components of DNA replication-dependent LexA cleavage Journal Article
| Authors: | Myka, K. K.; Marians, K. J. |
| Article Title: | Two components of DNA replication-dependent LexA cleavage |
| Abstract: | Induction of the SOS response, a cellular system triggered by DNA damage in bacteria, depends on DNA replication for the generation of the SOS signal, ssDNA. RecA binds to ssDNA, forming filaments that stimulate proteolytic cleavage of the LexA transcriptional repressor, allowing expression of > 40 gene products involved in DNA repair and cell cycle regulation. Here, using a DNA replication system reconstituted in vitro in tandem with a LexA cleavage assay, we studied LexA cleavage during DNA replication of both undamaged and base-damaged templates. Only a ssDNA-RecA filament supported LexA cleavage. Surprisingly, replication of an undamaged template supported levels of LexA cleavage like that induced by a template carrying two site-specific cyclobutane pyrimidine dimers. We found that two processes generate ssDNA that could support LexA cleavage. 1) During unperturbed replication, single-stranded regions formed because of stochastic uncoupling of the leading-strand DNA polymerase from the replication fork DNA helicase, and 2) on the damaged template, nascent leading-strand gaps were generated by replisome lesion skipping. The two pathways differed in that RecF stimulated LexA cleavage during replication of the damaged template, but not normal replication. RecF appears to facilitate RecA filament formation on the leading-strand ssDNA gaps generated by replisome lesion skipping. © 2020 Myka and Marians. |
| Keywords: | dna replication; enzyme degradation; e. coli; dna damage response; gene regulation; sos response; proteolytic cleavage; single-stranded dna; lexa; escherichia coli (e. coli) |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 295 |
| Issue: | 30 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2020-07-24 |
| Start Page: | 10368 |
| End Page: | 10379 |
| Language: | English |
| DOI: | 10.1074/jbc.RA120.014224 |
| PUBMED: | 32513870 |
| PROVIDER: | scopus |
| PMCID: | PMC7383369 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 1 September 2020 -- Source: Scopus |
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