An iron-sulfur cluster in the polymerase domain of yeast DNA polymerase ε

Rinku Jain; Eva S. Vanamee; Boris G. Dzikovski; Angeliki Buku; Robert Johnson; Louise Prakash; Satya Prakash; Aneel K. Aggarwal

doi:10.1016/j.jmb.2013.10.015

An iron-sulfur cluster in the polymerase domain of yeast DNA polymerase ε

Rinku Jain, Eva S. Vanamee, Boris G. Dzikovski, Angeliki Buku, Robert Johnson, Louise Prakash, Satya Prakash, Aneel K. Aggarwal

Biochemistry & Molecular Biology

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

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.

Original language	English (US)
Pages (from-to)	301-308
Number of pages	8
Journal	Journal of Molecular Biology
Volume	426
Issue number	2
DOIs	https://doi.org/10.1016/j.jmb.2013.10.015
State	Published - Jan 23 2014

Keywords

DNA polymerase
DNA repair
DNA replication
Fe-S cluster
oxidative stress

ASJC Scopus subject areas

Molecular Biology
Biophysics
Structural Biology

Access to Document

10.1016/j.jmb.2013.10.015

Cite this

@article{e6b8bda66ec742cb833518827015c196,

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 (Pol2ins) 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.",

keywords = "DNA polymerase, DNA repair, DNA replication, Fe-S cluster, oxidative stress",

author = "Rinku Jain and Vanamee, {Eva S.} and Dzikovski, {Boris G.} and Angeliki Buku and Robert Johnson and Louise Prakash and Satya Prakash and Aggarwal, {Aneel K.}",

note = "Funding Information: We thank the staff at Brookhaven National Laboratory (beamline X3B) for help with EXAFS data collection. The work was partially supported by grants CA138546 and CA107650 from the U.S. National Institutes of Health . The EPR studies were supported by National Institutes of Health grant P41GM103521 . ",

year = "2014",

month = jan,

day = "23",

doi = "10.1016/j.jmb.2013.10.015",

language = "English (US)",

volume = "426",

pages = "301--308",

journal = "Journal of Molecular Biology",

issn = "0022-2836",

publisher = "Academic Press",

number = "2",

}

TY - JOUR

T1 - An iron-sulfur cluster in the polymerase domain of yeast DNA polymerase ε

AU - Jain, Rinku

AU - Vanamee, Eva S.

AU - Dzikovski, Boris G.

AU - Buku, Angeliki

AU - Johnson, Robert

AU - Prakash, Louise

AU - Prakash, Satya

AU - Aggarwal, Aneel K.

N1 - Funding Information: We thank the staff at Brookhaven National Laboratory (beamline X3B) for help with EXAFS data collection. The work was partially supported by grants CA138546 and CA107650 from the U.S. National Institutes of Health . The EPR studies were supported by National Institutes of Health grant P41GM103521 .

PY - 2014/1/23

Y1 - 2014/1/23

N2 - 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 (Pol2ins) 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.

AB - 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 (Pol2ins) 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.

KW - DNA polymerase

KW - DNA repair

KW - DNA replication

KW - Fe-S cluster

KW - oxidative stress

UR - http://www.scopus.com/inward/record.url?scp=84891828842&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84891828842&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2013.10.015

DO - 10.1016/j.jmb.2013.10.015

M3 - Article

C2 - 24144619

AN - SCOPUS:84891828842

SN - 0022-2836

VL - 426

SP - 301

EP - 308

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 2

ER -

An iron-sulfur cluster in the polymerase domain of yeast DNA polymerase ε

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this