Mutations in the Escherichia coli Tus protein define a domain positioned close to the DNA in the Tus-Ter complex Journal Article
| Authors: | Skokotas, A.; Hiasa, H.; Marians, K. J.; O'Donnell, L.; Hill, T. M. |
| Article Title: | Mutations in the Escherichia coli Tus protein define a domain positioned close to the DNA in the Tus-Ter complex |
| Abstract: | A new genetic screen for mutations in the tus gene of Escherichia coli has been devised that selects for Tus proteins with altered ability to arrest DNA replication. We report here the catheterization of three such mutants: TusP42S, TusE49K, and TusH50Y. TusP42S and TusE49K arrest DNA replication in vivo at 36% of the efficiency of wild-type Tus, whereas TusH50Y functions at 78% efficiency. The loss of replication arrest activity did not correlate with changes in the stability of the Tus-TerB complexes formed by the mutant proteins. TusE49K formed a more stable protein-DNA complex than wild-type Tus (t( 1/2 ) of 178 versus 149 min, respectively) and TusP42 had a 9-min half- life, yet these two mutants showed identical efficiencies for replication arrest. When tested in vitro using a helicase assay or an oriC replication system, we observed a general, but imperfect, correlation between the in vivo and in vitro assays. Finally, the half-lives of the mutant protein-DNA complexes suggested that the domain of Tus where these mutations are located is positioned close to the DNA in the Tus-Ter complex. |
| Keywords: | gene mutation; dna-binding proteins; nonhuman; mutant protein; dna replication; protein domain; gene expression; protein binding; bacteria (microorganisms); bacterial protein; bacterial proteins; dna; molecular sequence data; kinetics; escherichia coli; plasmid; base sequence; helicase; genetic screening; mutagenesis; dna, bacterial; escherichia coli proteins; bacterial gene; dna protein complex; replication origin; priority journal; article |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 270 |
| Issue: | 52 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1995-12-29 |
| Start Page: | 30941 |
| End Page: | 30948 |
| Language: | English |
| DOI: | 10.1074/jbc.270.52.30941 |
| PUBMED: | 8537350 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 28 August 2018 -- Source: Scopus |
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