4RNO

Crystal structure of human polymerase eta extending an abasic site-dA pair by inserting dCTP opposite template G

4RNO の概要

• エントリーDOI: 10.2210/pdb4rno/pdb
• 関連するPDBエントリー: 4RNM 4RNN
• 分子名称: DNA polymerase eta, Nucleic acids Template: CATG(3DR)TGACGCT, Nucleic acids Primar: AGCGTCAA, ... (7 entities in total)
• 機能のキーワード: protein, dna, dna damage dna-directed dna polymerase, adenosine triphosphate, y-family polymerase, trans-lesion synthesis (tls), dna binding, abasic site lesion bypass, transferase-dna complex, transferase/dna
• 由来する生物種: Homo sapiens (human); 詳細
• 細胞内の位置: Nucleus : Q9Y253
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 55181.96

構造登録者

Patra, A.,Egli, M. (登録日: 2014-10-24, 公開日: 2015-02-18, 最終更新日: 2023-09-20)

主引用文献

Patra, A.,Zhang, Q.,Lei, L.,Su, Y.,Egli, M.,Guengerich, F.P.
Structural and Kinetic Analysis of Nucleoside Triphosphate Incorporation Opposite an Abasic Site by Human Translesion DNA Polymerase eta.
J.Biol.Chem., 290:8028-8038, 2015

PubMed Abstract: The most common lesion in DNA is an abasic site resulting from glycolytic cleavage of a base. In a number of cellular studies, abasic sites preferentially code for dATP insertion (the "A rule"). In some cases frameshifts are also common. X-ray structures with abasic sites in oligonucleotides have been reported for several microbial and human DNA polymerases (pols), e.g. Dpo4, RB69, KlenTaq, yeast pol ι, human (h) pol ι, and human pol β. We reported previously that hpol η is a major pol involved in abasic site bypass (Choi, J.-Y., Lim, S., Kim, E. J., Jo, A., and Guengerich, F. P. (2010 J. Mol. Biol. 404, 34-44). hpol η inserted all four dNTPs in steady-state and pre-steady-state assays, preferentially inserting A and G. In LC-MS analysis of primer-template pairs, A and G were inserted but little C or T was inserted. Frameshifts were observed when an appropriate pyrimidine was positioned 5' to the abasic site in the template. In x-ray structures of hpol η with a non-hydrolyzable analog of dATP or dGTP opposite an abasic site, H-bonding was observed between the phosphate 5' to the abasic site and water H-bonded to N1 and N6 of A and N1 and O6 of G nucleoside triphosphate analogs, offering an explanation for what appears to be a "purine rule." A structure was also obtained for an A inserted and bonded in the primer opposite the abasic site, but it did not pair with a 5' T in the template. We conclude that hpol η, a major copying enzyme with abasic sites, follows a purine rule, which can also lead to frameshifts. The phenomenon can be explained with H-bonds.

PubMed: 25666608
DOI: 10.1074/jbc.M115.637561

実験手法

X-RAY DIFFRACTION (2.82 Å)