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- Title
Eukaryotic DNA polymerases require an iron-sulfur cluster for the formation of active complexes.
- Authors
Netz, Daili J A; Stith, Carrie M; Stümpfig, Martin; Köpf, Gabriele; Vogel, Daniel; Genau, Heide M; Stodola, Joseph L; Lill, Roland; Burgers, Peter M J; Pierik, Antonio J
- Abstract
The eukaryotic replicative DNA polymerases (Pol ?, ? and ?) and the major DNA mutagenesis enzyme Pol ? contain two conserved cysteine-rich metal-binding motifs (CysA and CysB) in the C-terminal domain (CTD) of their catalytic subunits. Here we demonstrate by in vivo and in vitro approaches the presence of an essential [4Fe-4S] cluster in the CysB motif of all four yeast B-family DNA polymerases. Loss of the [4Fe-4S] cofactor by cysteine ligand mutagenesis in Pol3 destabilized the CTD and abrogated interaction with the Pol31 and Pol32 subunits. Reciprocally, overexpression of accessory subunits increased the amount of the CTD-bound Fe-S cluster. This implies an important physiological role of the Fe-S cluster in polymerase complex stabilization. Further, we demonstrate that the Zn-binding CysA motif is required for PCNA-mediated Pol ? processivity. Together, our findings show that the function of eukaryotic replicative DNA polymerases crucially depends on different metallocenters for accessory subunit recruitment and replisome stability.
- Subjects
DNA polymerases; EUKARYOTIC cells; IRON-sulfur proteins; MUTAGENESIS; CYSTEINE
- Publication
Nature Chemical Biology, 2012, Vol 8, Issue 1, p125
- ISSN
1552-4450
- Publication type
Article
- DOI
10.1038/nchembio.721