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

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)226-230
Number of pages5
JournalPhotochemistry and Photobiology
Volume69
Issue number2
DOIs
Publication statusPublished - Jan 1 1999
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Physical and Theoretical Chemistry

Cite this

Kobayashi, H., Sato, K., Komatsu, Y., Morioka, H., Stewart, J. D., Tsurimoto, T., & Ohtsuka, E. (1999). Effects of a High-Affinity Antibody Fragment on DNA Polymerase Reactions Near a (6-4) Photoproduct Site. Photochemistry and Photobiology, 69(2), 226-230. https://doi.org/10.1111/j.1751-1097.1999.tb03278.x