3UQ2

Crystal structure of the post-catalytic product complex of polymerase lambda with an rCMP inserted opposite a templating G and dAMP inserted opposite a templating T at the primer terminus.

3UQ2 の概要

• エントリーDOI: 10.2210/pdb3uq2/pdb
• 関連するPDBエントリー: 3MGH 3MGI 3UPQ 3UQ0 3UQ1
• 分子名称: DNA polymerase lambda, 5'-D(*CP*GP*GP*CP*TP*GP*TP*AP*CP*TP*G)-3', 5'-D(*CP*AP*GP*TP*AP)-R(P*CP*A)-3', ... (8 entities in total)
• 機能のキーワード: dna polymerase lambda, ribonucleotide incorporation, protein conformation, transferase, lyase-dna complex, lyase/dna
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Nucleus: Q9UGP5
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 43608.69

構造登録者

Gosavi, R.A.,Moon, A.F.,Kunkel, T.A.,Pedersen, L.C.,Bebenek, K. (登録日: 2011-11-18, 公開日: 2012-05-23, 最終更新日: 2023-09-13)

主引用文献

Gosavi, R.A.,Moon, A.F.,Kunkel, T.A.,Pedersen, L.C.,Bebenek, K.
The catalytic cycle for ribonucleotide incorporation by human DNA Pol lambda
Nucleic Acids Res., 40:7518-7527, 2012

PubMed Abstract: Although most DNA polymerases discriminate against ribonucleotide triphosphaets (rNTPs) during DNA synthesis, recent studies have shown that large numbers of ribonucleotides are incorporated into the eukaryotic nuclear genome. Here, we investigate how a DNA polymerase can stably incorporate an rNTP. The X-ray crystal structure of a variant of human DNA polymerase λ reveals that the rNTP occupies the nucleotide binding pocket without distortion of the active site, despite an unfavorable interaction between the 2'-O and Tyr505 backbone carbonyl. This indicates an energetically unstable binding state for the rNTP, stabilized by additional protein-nucleotide interactions. Supporting this idea is the 200-fold lower catalytic efficiency for rNTP relative to deoxyribonucleotide triphosphate (dNTP) incorporation, reflecting a higher apparent Km value for the rNTP. Furthermore, distortion observed in the structure of the post-catalytic product complex suggests that once the bond between the α- and β-phosphates of the rNTP is broken, the unfavorable binding state of the ribonucleotide cannot be maintained. Finally, structural and biochemical evaluation of dNTP insertion onto an ribonucleotide monophosphate (rNMP)-terminated primer indicates that a primer-terminal rNMP does not impede extension. The results are relevant to how ribonucleotides are incorporated into DNA in vivo, during replication and during repair, perhaps especially in non-proliferating cells when rNTP:dNTP ratios are high.

PubMed: 22584622
DOI: 10.1093/nar/gks413

実験手法

X-RAY DIFFRACTION (2.25 Å)