6VE5
X-ray structure of human REV7 in complex with Shieldin3 (residues 41-74)
Summary for 6VE5
| Entry DOI | 10.2210/pdb6ve5/pdb |
| Descriptor | Mitotic spindle assembly checkpoint protein MAD2B, Shieldin complex subunit 3, SULFATE ION, ... (4 entities in total) |
| Functional Keywords | 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 |
| Biological source | Homo sapiens (Human) More |
| Total number of polymer chains | 2 |
| Total formula weight | 28986.59 |
| Authors | Cui, G.,Botuyan, M.V.,Mer, G. (deposition date: 2019-12-28, release date: 2021-01-13, Last modification date: 2023-10-11) |
| Primary citation | 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 Cited by 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: 34174285DOI: 10.1016/j.jbc.2021.100912 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2 Å) |
Structure validation
Download full validation report
