Photoreactivation of DNA by an archaeal nucleoprotein Sso7d

摘要

Sso7d is a chromosomal protein of hyperthermophilic Archaea. The crystal structure of Sso7d-d(GTAAT(1)UAC)(2) has been clarified at high resolution, showing that the protein binds in the minor groove of DNA, causing a sharp kink of approximate to 60 degrees. Recently, we found that photoirradiation of Sso7d and 5-iodouracil-(U-1)-containing 5'-d(GTAAT(1)UAC)-3' efficiently induced the abstraction of hydrogen from the methyl group of T-5 at the kink. In the present study, we found that the photoreactivity of 5-bromouracil (U-Br)-containing 5'-d(GTAAT(Br)UAC)-3' was enhanced in the presence of Sso7d. Using hypoxanthine (I)-containing 5'-d(ITAAT(Br)UAC)-3', we demonstrated that electron transfer occurs efficiently from Sso7d to DNA. Product analysis showed that Trp-24 of Sso7d, located at the surface of the DNA, is consumed to produce N'-formylkynurenine during photoirradiation, indicating that Trp-24 acts as an electron source. To explore the possibility of electron transfer between Sso7d and other DNA substrates, we examined the photochemical repair of the thymine dimer 5'-d(GTAAT <> TAC)-3' by Sso7d. Sso7d effectively repaired 5'-d(GTAAT <> TAC)-3' to 5'-d(GTAATTAC)-3' under irradiation conditions. During this reaction, Trp-24 was not oxidized significantly, indicating that the anion radical of the repaired TT sequence is oxidized by the cation radical of Trp-24, and that a so-called "circular electron transfer" mechanism is operating in this system.