1997年4月
Substrate and mispairing properties of 5-formyl-2'-deoxyuridine 5'-triphosphate assessed by in vitro DNA polymerase reactions
NUCLEIC ACIDS RESEARCH
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- 巻
- 25
- 号
- 8
- 開始ページ
- 1570
- 終了ページ
- 1577
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1093/nar/25.8.1570
- 出版者・発行元
- OXFORD UNIV PRESS
5-Formyluracil (fU) is one of the thymine lesions produced by reactive oxygen radicals in DNA and its constituents. In this work, 5-formyl-2'-deoxyuridine 5'-triphosphate (fdUTP) was chemically synthesized and extensively purified by HPLC. The electron withdrawing 5-formyl group facilitated ionization of fU. Thus, pK(a) of the base unit of fdUTP was 8.6, significantly lower than that of parent thymine (pK(a) = 10.0 as dTMP). fdUTP efficiently replaced dTTP during DNA replication catalyzed by Escherichia coli DNA polymerase I (Klenow fragment), T7 DNA polymerase (3'-5' exonuclease free) and Taq DMA polymerase, fU-specific cleavage of the replication products by piperidine revealed that when incorporated as T, incorporation of fU was virtually uniform, suggesting minor sequence context effects on the incorporation frequency of fdUTP. fdUTP also replaced dCTP, but with much Bower efficiency than that for dTTP, The substitution efficiency for dCTP increased with increasing pH from 7.2 to 9.0, The parallel correlation between ionization of the base unit of fdUTP (pK(a) = 8.6) and the Substitution efficiency for dCTP strongly suggests that the base-ionized form of dFdUTP is involved in mispairing with template G. These data indicate that fU can be specifically introduced into DNA as unique lesions by in vitro DNA polymerase reactions. in addition, fU is potentially mutagenic since this lesion is much more prone to form mispairing with G than parent thymine.
- リンク情報
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- DOI
- https://doi.org/10.1093/nar/25.8.1570
- CiNii Articles
- http://ci.nii.ac.jp/naid/30018406052
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/9092664
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:A1997WV07800015&DestApp=WOS_CPL
- ID情報
-
- DOI : 10.1093/nar/25.8.1570
- ISSN : 0305-1048
- CiNii Articles ID : 30018406052
- PubMed ID : 9092664
- Web of Science ID : WOS:A1997WV07800015