論文 - 詳細
| RRC ID | 85521 |
|---|---|
| 著者 | Pham TM, Tan KW, Sakumura Y, Okumura K, Maki H, Akiyama MT. |
| タイトル | A single-molecule approach to DNA replication in Escherichia coli cells demonstrated that DNA polymerase III is a major determinant of fork speed. |
| ジャーナル | Mol Microbiol |
| Abstract |
The replisome catalyses DNA synthesis at a DNA replication fork. The molecular behaviour of the individual replisomes, and therefore the dynamics of replication fork movements, in growing Escherichia coli cells remains unknown. DNA combing enables a single-molecule approach to measuring the speed of replication fork progression in cells pulse-labelled with thymidine analogues. We constructed a new thymidine-requiring strain, eCOMB (E. coli for combing), that rapidly and sufficiently incorporates the analogues into newly synthesized DNA chains for the DNA-combing method. In combing experiments with eCOMB, we found the speed of most replication forks in the cells to be within the narrow range of 550-750 nt s(-1) and the average speed to be 653 ± 9 nt s(-1) (± SEM). We also found the average speed of the replication fork to be only 264 ± 9 nt s(-1) in a dnaE173-eCOMB strain producing a mutant-type of the replicative DNA polymerase III (Pol III) with a chain elongation rate (300 nt s(-1) ) much lower than that of the wild-type Pol III (900 nt s(-1) ). This indicates that the speed of chain elongation by Pol III is a major determinant of replication fork speed in E. coli cells. |
| 巻・号 | 90(3) |
| ページ | 584-96 |
| 公開日 | 2013-11-1 |
| DOI | 10.1111/mmi.12386 |
| PMID | 23998701 |
| MeSH | Bromodeoxyuridine Chromosomes, Bacterial DNA Polymerase III / genetics DNA Polymerase III / metabolism* DNA Replication* DNA, Bacterial / biosynthesis* DNA, Bacterial / metabolism Escherichia coli / genetics Escherichia coli / growth & development* Escherichia coli / metabolism Escherichia coli Proteins / genetics Escherichia coli Proteins / metabolism Mutation Thymidine / analogs & derivatives |
| IF | 3.418 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 原核生物(大腸菌) | ME10091 |