title: An iron-sulfur cluster in the polymerase domain of yeast DNA polymerase ε.
abstract: DNA polymerase ε (Polε) is a multi-subunit polymerase that contributes to genomic stability via its roles in leading strand replication and the repair of damaged DNA. Polε from Saccharomyces cerevisiae is composed of four subunits--Pol2, Dpb2, Dpb3, and Dpb4. Here, we report the presence of a [Fe-S] cluster directly within the active polymerase domain of Pol2 (residues 1-1187). We show that binding of the [Fe-S] cluster is mediated by cysteines in an insertion (Pol2(ins)) that is conserved in Pol2 orthologs but is absent in the polymerase domains of Polα, Polδ, and Polζ. We also show that the [Fe-S] cluster is required for Pol2 polymerase activity but not for its exonuclease activity. Collectively, our work suggests that Polε is perhaps more sensitive than other DNA polymerases to changes in oxidative stress in eukaryotic cells.
authors.affiliation: Department of Structural and Chemical Biology, Mount Sinai School of Medicine, Box 1677, 1425 Madison Avenue, New York, NY 10029, USA.
authors.forename: Rinku
authors.initials: R
authors.lastname: Jain
authors.affiliation: Department of Structural and Chemical Biology, Mount Sinai School of Medicine, Box 1677, 1425 Madison Avenue, New York, NY 10029, USA.
authors.forename: Eva S
authors.initials: ES
authors.lastname: Vanamee
authors.affiliation: Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Boris G
authors.initials: BG
authors.lastname: Dzikovski
authors.affiliation: Department of Structural and Chemical Biology, Mount Sinai School of Medicine, Box 1677, 1425 Madison Avenue, New York, NY 10029, USA.
authors.forename: Angeliki
authors.initials: A
authors.lastname: Buku
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77755-1061, USA.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Johnson
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77755-1061, USA.
authors.forename: Louise
authors.initials: L
authors.lastname: Prakash
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77755-1061, USA.
authors.forename: Satya
authors.initials: S
authors.lastname: Prakash
authors.affiliation: Department of Structural and Chemical Biology, Mount Sinai School of Medicine, Box 1677, 1425 Madison Avenue, New York, NY 10029, USA. Electronic address: Aneel.Aggarwal@mssm.edu.
authors.forename: Aneel K
authors.initials: AK
authors.lastname: Aggarwal
mesh_terms: D004257:DNA Polymerase II / Q000737:chemistry* / Q000378:metabolism*; D007501:Iron / Q000032:analysis*; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism*; D018384:Oxidative Stress; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013455:Sulfur / Q000032:analysis*
chemical_list: D007506:Iron-Sulfur Proteins; D013455:Sulfur; D007501:Iron; D004257:DNA Polymerase II
keywords: DNA polymerase; DNA repair; DNA replication; EXAFS; FT-EXAFS; Fe-S cluster; Fourier transform EXAFS; ICP; extended X-ray absorption fine structure; inductively coupled plasma; oxidative stress