6VE5

X-ray structure of human REV7 in complex with Shieldin3 (residues 41-74)

6VE5 の概要

• エントリーDOI: 10.2210/pdb6ve5/pdb
• 分子名称: Mitotic spindle assembly checkpoint protein MAD2B, Shieldin complex subunit 3, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: dna damage response, dna double-strand break repair, dna end resection, homologous recombination, non-homologous end joining, shieldin3, shld3, rinn1, rev7, 53bp1, rif1, protein binding
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 28986.59

構造登録者

Cui, G.,Botuyan, M.V.,Mer, G. (登録日: 2019-12-28, 公開日: 2021-01-13, 最終更新日: 2023-10-11)

主引用文献

Du Truong, C.,Craig, T.A.,Cui, G.,Botuyan, M.V.,Serkasevich, R.A.,Chan, K.Y.,Mer, G.,Chiu, P.L.,Kumar, R.
Cryo-EM reveals conformational flexibility in apo DNA polymerase zeta.
J.Biol.Chem., 297:100912-100912, 2021

PubMed Abstract: The translesion synthesis (TLS) DNA polymerases Rev1 and Polζ function together in DNA lesion bypass during DNA replication, acting as nucleotide inserter and extender polymerases, respectively. While the structural characterization of the Saccharomyces cerevisiae Polζ in its DNA-bound state has illuminated how this enzyme synthesizes DNA, a mechanistic understanding of TLS also requires probing conformational changes associated with DNA- and Rev1 binding. Here, we used single-particle cryo-electron microscopy to determine the structure of the apo Polζ holoenzyme. We show that compared with its DNA-bound state, apo Polζ displays enhanced flexibility that correlates with concerted motions associated with expansion of the Polζ DNA-binding channel upon DNA binding. We also identified a lysine residue that obstructs the DNA-binding channel in apo Polζ, suggesting a gating mechanism. The Polζ subunit Rev7 is a hub protein that directly binds Rev1 and is a component of several other protein complexes such as the shieldin DNA double-strand break repair complex. We analyzed the molecular interactions of budding yeast Rev7 in the context of Polζ and those of human Rev7 in the context of shieldin using a crystal structure of Rev7 bound to a fragment of the shieldin-3 protein. Overall, our study provides new insights into Polζ mechanism of action and the manner in which Rev7 recognizes partner proteins.

PubMed: 34174285
DOI: 10.1016/j.jbc.2021.100912

実験手法

X-RAY DIFFRACTION (2 Å)