6H1V

The crystal structure of Pol2CORE in complex with DNA and an incoming nucleotide, carrying an Fe-S cluster

6H1V の概要

• エントリーDOI: 10.2210/pdb6h1v/pdb
• 関連するPDBエントリー: 6gzg
• 分子名称: DNA polymerase epsilon catalytic subunit A, DNA (5'-D(P*TP*AP*AP*CP*CP*GP*CP*GP*TP*TP*(DOC))-3'), DNA (5'-D(P*AP*AP*TP*TP*GP*AP*AP*CP*GP*CP*GP*GP*TP*TP*A)-3'), ... (7 entities in total)
• 機能のキーワード: pol epsilon, m644g, dna, complex, dna binding protein, fe-s
• 由来する生物種: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 145787.77

構造登録者

Parkash, V.,Johansson, E. (登録日: 2018-07-12, 公開日: 2019-04-24, 最終更新日: 2024-01-17)

主引用文献

Ter Beek, J.,Parkash, V.,Bylund, G.O.,Osterman, P.,Sauer-Eriksson, A.E.,Johansson, E.
Structural evidence for an essential Fe-S cluster in the catalytic core domain of DNA polymerase ε.
Nucleic Acids Res., 47:5712-5722, 2019

PubMed Abstract: DNA polymerase ϵ (Pol ϵ), the major leading-strand DNA polymerase in eukaryotes, has a catalytic subunit (Pol2) and three non-catalytic subunits. The N-terminal half of Pol2 (Pol2CORE) exhibits both polymerase and exonuclease activity. It has been suggested that both the non-catalytic C-terminal domain of Pol2 (with the two cysteine motifs CysA and CysB) and Pol2CORE (with the CysX cysteine motif) are likely to coordinate an Fe-S cluster. Here, we present two new crystal structures of Pol2CORE with an Fe-S cluster bound to the CysX motif, supported by an anomalous signal at that position. Furthermore we show that purified four-subunit Pol ϵ, Pol ϵ CysAMUT (C2111S/C2133S), and Pol ϵ CysBMUT (C2167S/C2181S) all have an Fe-S cluster that is not present in Pol ϵ CysXMUT (C665S/C668S). Pol ϵ CysAMUT and Pol ϵ CysBMUT behave similarly to wild-type Pol ϵ in in vitro assays, but Pol ϵ CysXMUT has severely compromised DNA polymerase activity that is not the result of an excessive exonuclease activity. Tetrad analyses show that haploid yeast strains carrying CysXMUT are inviable. In conclusion, Pol ϵ has a single Fe-S cluster bound at the base of the P-domain, and this Fe-S cluster is essential for cell viability and polymerase activity.

PubMed: 30968138
DOI: 10.1093/nar/gkz248

実験手法

X-RAY DIFFRACTION (2.7 Å)