Purification and characterization of DnaC810, a primosomal protein capable of bypassing PriA function Journal Article
| Authors: | Xu, L.; Marians, K. J. |
| Article Title: | Purification and characterization of DnaC810, a primosomal protein capable of bypassing PriA function |
| Abstract: | Escherichia coli strains lacking PriA are severely compromised in their ability to repair UV-damaged DNA and to perform homologous recombination. These phenotypes arise because of a lack of PriA-directed replication fork assembly at recombination intermediates such as D-loops. Naturally arising suppressor mutations in dnaC restore strains carrying the priA2::kan null allele to wild-type function. We have cloned one such gene, dnaC810, and overexpressed, purified, and characterized the DnaC810 protein. DnaC810 can support a PriA-independent synthesis of φX174 complementary strand DNA. This can be attributed to its ability, unlike wild-type DnaC, to catalyze a SSB- insensitive general priming reaction with DnaB and DnaG on any SSB-coated single-stranded DNA. Gel mobility shift analysis revealed that DnaC810 could load DnaB directly to SSB-coated single-stranded DNA as well as to D loop DNA. This explains the ability of DnaC810 to bypass the requirement for PriA, PriB, PriC, and DnaT during replication fork assembly at recombination intermediates. |
| Keywords: | mutation; dna-binding proteins; nonhuman; dna replication; dna recombination; dna damage; dna repair; gene overexpression; protein binding; bacteria (microorganisms); bacterial protein; bacterial proteins; escherichia coli; recombinant proteins; dna, single-stranded; dna determination; dna helicases; replication protein a; escherichia coli proteins; dna purification; prokaryota; suppression, genetic; dnab helicases; bacteriophage phi x 174; priority journal; article |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 275 |
| Issue: | 11 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2000-03-17 |
| Start Page: | 8196 |
| End Page: | 8205 |
| Language: | English |
| DOI: | 10.1074/jbc.275.11.8196 |
| PUBMED: | 10713144 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
