論文 - 詳細
| RRC ID | 35536 |
|---|---|
| 著者 | Conte E, Vincelli G, Schaaper RM, Bressanin D, Stefan A, Dal Piaz F, Hochkoeppler A. |
| タイトル | Stabilization of the Escherichia coli DNA polymerase III ε subunit by the θ subunit favors in vivo assembly of the Pol III catalytic core. |
| ジャーナル | Arch Biochem Biophys |
| Abstract |
Escherichia coli DNA polymerase III holoenzyme (HE) contains a core polymerase consisting of three subunits: α (polymerase), ε (3'-5' exonuclease), and θ. Genetic experiments suggested that θ subunit stabilizes the intrinsically labile ε subunit and, furthermore, that θ might affect the cellular amounts of Pol III core and HE. Here, we provide biochemical evidence supporting this model by analyzing the amounts of the relevant proteins. First, we show that a ΔholE strain (lacking θ subunit) displays reduced amounts of free ε. We also demonstrate the existence of a dimer of ε, which may be involved in the stabilization of the protein. Second, θ, when overexpressed, dissociates the ε dimer and significantly increases the amount of Pol III core. The stability of ε also depends on cellular chaperones, including DnaK. Here, we report that: (i) temperature shift-up of ΔdnaK strains leads to rapid depletion of ε, and (ii) overproduction of θ overcomes both the depletion of ε and the temperature sensitivity of the strain. Overall, our data suggest that ε is a critical factor in the assembly of Pol III core, and that this is role is strongly influenced by the θ subunit through its prevention of ε degradation. |
| 巻・号 | 523(2) |
| ページ | 135-43 |
| 公開日 | 2012-7-15 |
| DOI | 10.1016/j.abb.2012.04.013 |
| PII | S0003-9861(12)00145-2 |
| PMID | 22546509 |
| PMC | PMC3441146 |
| MeSH | Catalytic Domain* DNA Polymerase III / chemistry* DNA Polymerase III / deficiency DNA Polymerase III / metabolism* Enzyme Stability Escherichia coli / enzymology* Escherichia coli / genetics Escherichia coli Proteins / chemistry* Escherichia coli Proteins / metabolism* Gene Expression Regulation, Bacterial HSP70 Heat-Shock Proteins / metabolism Protein Multimerization Protein Structure, Quaternary |
| IF | 3.391 |
| 引用数 | 8 |
| WOS 分野 | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| 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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| 各媒体での言及数の合計 | 0 |
| 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) | JW0013-KC? JW1831-KC? |