Effects of a High-Affinity Antibody Fragment on DNA Polymerase Reactions Near a (6-4) Photoproduct Site

Hiroyuki Kobayashi, Kousuke Sato, Yasuo Komatsu, Hiroshi Morioka, Jon D. Stewart, Toshiki Tsurimoto, Eiko Ohtsuka

研究成果: Contribution to journalArticle査読

6 被引用数 (Scopus)

抄録

Pyrimidine (6-4) pyrimidone photodimers are major photoproducts that have mutagenic and carcinogenic consequences. One major reason for these biological effects of (6-4) photoproducts may be base mispairing/DNA replication errors due to hydrogen bonding to bases opposite these damaged sites. We synthesized a modified 41-mer DNA containing a (6-4) photoproduct using a preformed building block, then employed it as a template for primer extension reactions catalyzed by Klenow fragment and DNA polymerases α, β and δ (pol α, pol β and pol δ). None of these DNA polymerases were able to bypass the (6-4) photoproduct and elongation terminated at or near the 3′-pyrimidone of the photoproduct, depending on the dNTP concentration. When a single-chain Fv (scFv) with high affinity for the (6-4) photoproduct was included in the polymerization reaction, DNA synthesis was inhibited at base positions four, six, eight or eight nucleotides prior to the 3′-pyrimidone by Klenow fragment, pol α, pol β or pol δ, respectively. These results suggest that the scFv can bind to the template DNA containing a (6-4) photoproduct and inhibit extension reactions by polymerases.

本文言語英語
ページ(範囲)226-230
ページ数5
ジャーナルPhotochemistry and Photobiology
69
2
DOI
出版ステータス出版済み - 1 1 1999
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生化学
  • 物理化学および理論化学

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