6JUQ

mutant PolIV-DNA incoming nucleotide complex 2

6JUQ の概要

• エントリーDOI: 10.2210/pdb6juq/pdb
• 分子名称: DNA polymerase IV, DNA (5'-D(P*CP*TP*GP*GP*GP*GP*TP*CP*CP*TP*AP*GP*GP*AP*CP*CP*C)-3'), DNA (5'-D(P*GP*GP*GP*TP*CP*CP*TP*AP*GP*GP*AP*CP*CP*C)-3'), ... (8 entities in total)
• 機能のキーワード: dna polymerase, replication, replication-dna complex, replication/dna
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 97119.40

構造登録者

Nair, D.T.,Johnson, M.K. (登録日: 2019-04-15, 公開日: 2020-02-19, 最終更新日: 2023-11-22)

主引用文献

Johnson, M.K.,Kottur, J.,Nair, D.T.
A polar filter in DNA polymerases prevents ribonucleotide incorporation.
Nucleic Acids Res., 47:10693-10705, 2019

PubMed Abstract: The presence of ribonucleotides in DNA can lead to genomic instability and cellular lethality. To prevent adventitious rNTP incorporation, the majority of the DNA polymerases (dPols) possess a steric filter. The dPol named MsDpo4 (Mycobacterium smegmatis) naturally lacks this steric filter and hence is capable of rNTP addition. The introduction of the steric filter in MsDpo4 did not result in complete abrogation of the ability of this enzyme to incorporate ribonucleotides. In comparison, DNA polymerase IV (PolIV) from Escherichia coli exhibited stringent selection for deoxyribonucleotides. A comparison of MsDpo4 and PolIV led to the discovery of an additional polar filter responsible for sugar selectivity. Thr43 represents the filter in PolIV and this residue forms interactions with the incoming nucleotide to draw it closer to the enzyme surface. As a result, the 2'-OH in rNTPs will clash with the enzyme surface, and therefore ribonucleotides cannot be accommodated in the active site in a conformation compatible with productive catalysis. The substitution of the equivalent residue in MsDpo4-Cys47, with Thr led to a drastic reduction in the ability of the mycobacterial enzyme to incorporate rNTPs. Overall, our studies evince that the polar filter serves to prevent ribonucleotide incorporation by dPols.

PubMed: 31544946
DOI: 10.1093/nar/gkz792

実験手法

X-RAY DIFFRACTION (2.74 Å)