5L1J

Crystal Structure of Human DNA Polymerase Eta Inserting dTMPNPP Opposite O6-Methyl-2'-deoxyguanosine

5L1J の概要

• エントリーDOI: 10.2210/pdb5l1j/pdb
• 関連するPDBエントリー: 5L1I 5L1K 5L1L
• 分子名称: DNA polymerase eta, DNA (5'-D(*CP*AP*TP*(6OG)P*AP*TP*GP*AP*CP*GP*CP*T)-3'), DNA (5'-D(*AP*GP*CP*GP*TP*CP*AP*T)-3'), ... (7 entities in total)
• 機能のキーワード: catalytic domain, dna damage, dna polymerase, lesion bypass, o6-methyl-2'-deoxyguanosine, y-family polymerase, translesion dna synthesis (tls), dna binding protein., 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: 55342.64

構造登録者

Patra, A.,Egli, M. (登録日: 2016-07-29, 公開日: 2016-10-05, 最終更新日: 2023-10-04)

主引用文献

Patra, A.,Zhang, Q.,Guengerich, F.P.,Egli, M.
Mechanisms of Insertion of dCTP and dTTP Opposite the DNA Lesion O6-Methyl-2'-deoxyguanosine by Human DNA Polymerase eta.
J.Biol.Chem., 291:24304-24313, 2016

PubMed Abstract: O-Methyl-2'-deoxyguanosine (O-MeG) is a ubiquitous DNA lesion, formed not only by xenobiotic carcinogens but also by the endogenous methylating agent S-adenosylmethionine. It can introduce mutations during DNA replication, with different DNA polymerases displaying different ratios of correct or incorrect incorporation opposite this nucleoside. Of the "translesion" Y-family human DNA polymerases (hpols), hpol η is most efficient in incorporating equal numbers of correct and incorrect C and T bases. However, the mechanistic basis for this specific yet indiscriminate activity is not known. To explore this question, we report biochemical and structural analysis of the catalytic core of hpol η. Activity assays showed the truncated form displayed similar misincorporation properties as the full-length enzyme, incorporating C and T equally and extending from both. X-ray crystal structures of both dC and dT paired with O-MeG were solved in both insertion and extension modes. The structures revealed a Watson-Crick-like pairing between O-MeG and 2"-deoxythymidine-5"-[(α, β)-imido]triphosphate (approximating dT) at both the insertion and extension stages with formation of two H-bonds. Conversely, both the structures with O- MeG opposite dCTP and dC display sheared configuration of base pairs but to different degrees, with formation of two bifurcated H-bonds and two single H-bonds in the structures trapped in the insertion and extension states, respectively. The structural data are consistent with the observed tendency of hpol η to insert both dC and dT opposite the O-MeG lesion with similar efficiencies. Comparison of the hpol η active site configurations with either O-MeG:dC or O-MeG:dT bound compared with the corresponding situations in structures of complexes of Sulfolobus solfataricus Dpo4, a bypass pol that favors C relative to T by a factor of ∼4, helps rationalize the more error-prone synthesis opposite the lesion by hpol η.

PubMed: 27694439
DOI: 10.1074/jbc.M116.755462

実験手法

X-RAY DIFFRACTION (1.94 Å)