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-Structure paper
タイトル | Coordinated DNA polymerization by Polγ and the region of LonP1 regulated proteolysis. |
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ジャーナル・号・ページ | Nucleic Acids Res, Vol. 52, Issue 13, Page 7863-7875, Year 2024 |
掲載日 | 2024年7月22日 |
著者 | Amanda A Riccio / Asia J Brannon / Juno M Krahn / Jonathan Bouvette / Jason G Williams / Mario J Borgnia / William C Copeland / |
PubMed 要旨 | The replicative mitochondrial DNA polymerase, Polγ, and its protein regulation are essential for the integrity of the mitochondrial genome. The intricacies of Polγ regulation and its interactions ...The replicative mitochondrial DNA polymerase, Polγ, and its protein regulation are essential for the integrity of the mitochondrial genome. The intricacies of Polγ regulation and its interactions with regulatory proteins, which are essential for fine-tuning polymerase function, remain poorly understood. Misregulation of the Polγ heterotrimer, consisting of (i) PolG, the polymerase catalytic subunit and (ii) PolG2, the accessory subunit, ultimately results in mitochondrial diseases. Here, we used single particle cryo-electron microscopy to resolve the structure of PolG in its apoprotein state and we captured Polγ at three intermediates within the catalytic cycle: DNA bound, engaged, and an active polymerization state. Chemical crosslinking mass spectrometry, and site-directed mutagenesis uncovered the region of LonP1 engagement of PolG, which promoted proteolysis and regulation of PolG protein levels. PolG2 clinical variants, which disrupted a stable Polγ complex, led to enhanced LonP1-mediated PolG degradation. Overall, this insight into Polγ aids in an understanding of mitochondrial DNA replication and characterizes how machinery of the replication fork may be targeted for proteolytic degradation when improperly functioning. |
リンク | Nucleic Acids Res / PubMed:38932681 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.0 - 4.2 Å |
構造データ | EMDB-42979, PDB-8v54: EMDB-42980, PDB-8v55: EMDB-42982, PDB-8v5d: EMDB-42984, PDB-8v5r: |
化合物 | ChemComp-MG: ChemComp-D3T: |
由来 |
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キーワード | DNA BINDING PROTEIN/DNA / Mitochondria DNA polymerase / DNA polymerase gamma / Mitochondrial DNA replication / DNA binding protein / DNA BINDING PROTEIN-DNA complex / DNA polymerase / DNA polymerase catalytic subunit |