Eukaryotic DNA polymerases require an iron-sulfur cluster for the formation of active complexes

Daili J.A. Netz, Carrie M. Stith, Martin Stümpfig, Gabriele Köpf, Daniel Vogel, Heide M. Genau, Joseph L. Stodola, Roland Lill, Peter M.J. Burgers, Antonio J. Pierik

Research output: Contribution to journalArticlepeer-review

302 Scopus citations


The eukaryotic replicative DNA polymerases (Pol α, δ and ε) and the major DNA mutagenesis enzyme Pol ζ 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 δ 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.

Original languageEnglish
Pages (from-to)125-132
Number of pages8
JournalNature Chemical Biology
Issue number1
StatePublished - Jan 2012


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