Temperature-sensitive photoreactivation of cyclobutane thymine dimer in soybean

Ayumi Yamamoto, Najrana Tanbir, Tokuhisa Hirouchi, Mika Teranishi, Jun Hidema, Hiroshi Morioka, Kazuo Yamamoto

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

UV radiation induces the formation of two classes of photoproducts in DNA, the cyclobutane pyrimidine dimer (CPD) and the pyrimidine 6-4 pyrimidone photoproduct. CPDs in plants are repaired by class II CPD photolyase via a UV-A/blue light-dependent mechanism. The genes for the class II CPD photolyase have been cloned from higher plants such as Arabidopsis, Cucumis sativus (cucumber), Oryza sativa (rice) and Spinacia oleracea (spinach). Flavin adenine dinucleotide (FAD) has been identified as a cofactor. Here we report the isolation and characterization of the CPD photolyase cDNA from soybean (Glycin max). The sequence of amino acids predicted from the cDNA sequence was highly homologous to sequences of higher plant class II CPD photolyases. When the cDNA was expressed in a photolyasedeficient Escherichia coli, photoreactivation activity was partially restored by illumination with a fluorescent light. The purified enzyme showed CPD binding and light-dependent photoreactivation activities in vitro. When soybean CPD photolyase was heat-treated in vitro from 25°C to 45°C for 3 min, thymine dimer-binding activity and photoreactivation activity were decreased, and FAD was released from the enzyme. On the other hand, when the enzyme-CPD complex was heat-treated, photoreactivation activity was stable. We argue that FAD in the soybean CPD photolyase is labile for temperature, but once the enzyme-CPD complex has formed, FAD becomes tightly bound to the enzyme or complex.

Original languageEnglish
Pages (from-to)189-196
Number of pages8
JournalJournal of radiation research
Volume49
Issue number2
DOIs
Publication statusPublished - 2008

Keywords

  • FAD
  • Photolyase
  • Soybean (Glycin max)
  • Thermostability

ASJC Scopus subject areas

  • Radiation
  • Radiology Nuclear Medicine and imaging
  • Health, Toxicology and Mutagenesis

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