PT  - JOURNAL ARTICLE
AU  - Stanislaw K. Jozwiakowski
AU  - Sandra Kummer
AU  - Kerstin Gari
TI  - Human DNA polymerase delta requires an iron–sulfur cluster for high-fidelity DNA synthesis
AID  - 10.26508/lsa.201900321
DP  - 2019 Aug 01
TA  - Life Science Alliance
PG  - e201900321
VI  - 2
IP  - 4
4099  - https://www.life-science-alliance.org/content/2/4/e201900321.short
4100  - https://www.life-science-alliance.org/content/2/4/e201900321.full
SO  - Life Sci. Alliance2019 Aug 01; 2
AB  - Replication of eukaryotic genomes relies on the family B DNA polymerases Pol α, Pol δ, and Pol ε. All of these enzymes coordinate an iron–sulfur (FeS) cluster, but the function of this cofactor has remained largely unclear. Here, we show that the FeS cluster in the catalytic subunit of human Pol δ is coordinated by four invariant cysteines of the C-terminal CysB motif. FeS cluster loss causes a partial destabilisation of the four-subunit enzyme, a defect in double-stranded DNA binding, and compromised polymerase and exonuclease activities. Importantly, complex stability, DNA binding, and enzymatic activities are restored in the presence of proliferating cell nuclear antigen. We further show that also more subtle changes to the FeS cluster-binding pocket that do not abolish FeS cluster binding can have repercussions on the distant exonuclease domain and render the enzyme error-prone. Our data hence suggest that the FeS cluster in human Pol δ is an important co-factor that despite its C-terminal location has an impact on both DNA polymerase and exonuclease activities, and can influence the fidelity of DNA synthesis.