7NV0

Human Pol Kappa holoenzyme with wt PCNA

7NV0 の概要

• エントリーDOI: 10.2210/pdb7nv0/pdb
• 関連するPDBエントリー: 7NV1
• EMDBエントリー: 12601 12602
• 分子名称: DNA polymerase kappa, Proliferating cell nuclear antigen, DNA Primer, ... (5 entities in total)
• 機能のキーワード: translesion synthesis, tls, replication
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 206042.57

構造登録者

Lancey, C.,De Biasio, A.,Hamdan, S.M. (登録日: 2021-03-15, 公開日: 2021-11-10, 最終更新日: 2024-07-10)

主引用文献

Lancey, C.,Tehseen, M.,Bakshi, S.,Percival, M.,Takahashi, M.,Sobhy, M.A.,Raducanu, V.S.,Blair, K.,Muskett, F.W.,Ragan, T.J.,Crehuet, R.,Hamdan, S.M.,De Biasio, A.
Cryo-EM structure of human Pol kappa bound to DNA and mono-ubiquitylated PCNA.
Nat Commun, 12:6095-6095, 2021

PubMed Abstract: Y-family DNA polymerase κ (Pol κ) can replicate damaged DNA templates to rescue stalled replication forks. Access of Pol κ to DNA damage sites is facilitated by its interaction with the processivity clamp PCNA and is regulated by PCNA mono-ubiquitylation. Here, we present cryo-EM reconstructions of human Pol κ bound to DNA, an incoming nucleotide, and wild type or mono-ubiquitylated PCNA (Ub-PCNA). In both reconstructions, the internal PIP-box adjacent to the Pol κ Polymerase-Associated Domain (PAD) docks the catalytic core to one PCNA protomer in an angled orientation, bending the DNA exiting the Pol κ active site through PCNA, while Pol κ C-terminal domain containing two Ubiquitin Binding Zinc Fingers (UBZs) is invisible, in agreement with disorder predictions. The ubiquitin moieties are partly flexible and extend radially away from PCNA, with the ubiquitin at the Pol κ-bound protomer appearing more rigid. Activity assays suggest that, when the internal PIP-box interaction is lost, Pol κ is retained on DNA by a secondary interaction between the UBZs and the ubiquitins flexibly conjugated to PCNA. Our data provide a structural basis for the recruitment of a Y-family TLS polymerase to sites of DNA damage.

PubMed: 34667155
DOI: 10.1038/s41467-021-26251-6

実験手法

ELECTRON MICROSCOPY (3.4 Å)