2M2I

NMR solution structure of BRCT domain of yeast REV1

2M2I の概要

• エントリーDOI: 10.2210/pdb2m2i/pdb
• NMR情報: BMRB: 18916
• 分子名称: DNA repair protein REV1 (1 entity in total)
• 機能のキーワード: brct, rev1, transferase
• 由来する生物種: Saccharomyces cerevisiae (Baker's yeast)
• 細胞内の位置: Nucleus: P12689
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 10792.60

構造登録者

Pustovalova, Y.,Maciejewski, M.W.,Korzhnev, D.M. (登録日: 2012-12-21, 公開日: 2013-01-16, 最終更新日: 2024-05-15)

主引用文献

Pustovalova, Y.,Maciejewski, M.W.,Korzhnev, D.M.
NMR mapping of PCNA interaction with translesion synthesis DNA polymerase Rev1 mediated by Rev1-BRCT domain.
J.Mol.Biol., 425:3091-3105, 2013

PubMed Abstract: Rev1 is a Y-family translesion synthesis (TLS) DNA polymerase involved in bypass replication across sites of DNA damage and postreplicational gap filling. In the process of TLS, high-fidelity replicative DNA polymerases stalled by DNA damage are replaced by error-prone TLS enzymes responsible for the majority of mutagenesis in eukaryotic cells. The polymerase exchange that gains low-fidelity TLS polymerases access to DNA is mediated by their interactions with proliferating cell nuclear antigen (PCNA). Rev1 stands alone from other Y-family TLS enzymes since it lacks the consensus PCNA-interacting protein box (PIP-box) motif, instead utilizing other modular domains for PCNA binding. Here we report solution NMR structure of an 11-kDa BRCA1 C-terminus (BRCT) domain from Saccharomyces cerevisiae Rev1 and demonstrate with the use of transverse relaxation optimized spectroscopy (TROSY) NMR methods that Rev1-BRCT domain directly interacts with an 87-kDa PCNA in solution. The domain adopts α/β fold (β1-α1-β2-β3-α2-β4-α3-α4) typical for BRCT domain superfamily. PCNA-binding interface of the Rev1-BRCT domain comprises conserved residues of the outer surface of the α1-helix and the α1-β1, β2-β3 and β3-α2 loops. On the other hand, Rev1-BRCT binds to the inter-domain region of PCNA that overlaps with the binding site for the PIP-box motif. Furthermore, Rev1-BRCT domain bound to PCNA can be displaced by increasing amounts of the PIP-box peptide from TLS DNA polymerase polη, suggesting that Rev1-BRCT and polη PIP-box interactions with the same PCNA monomer are mutually exclusive. These results provide structural insights into PCNA recognition by TLS DNA polymerases that help better understand TLS regulation in eukaryotes.

PubMed: 23747975
DOI: 10.1016/j.jmb.2013.05.029

実験手法

SOLUTION NMR