Inhibition of restriction enzyme's DNA sequence recognition by PUVA treatment.
Applying various restriction enzymes on a specially designed 1.5 kb DNA fragment revealed that the inhibitory effects of psoralens + UVA irradiation (PUVA) treatment on restriction endonuclease activities are caused by recognition inhibition. In this study restriction enzymes that have a 5'-TpA sequence at the cleaving site (KpnI, XbaI, PmeI and DraI), and the noncleaving site (PacI) in recognition sites, or have two 5'-TpA sequences at the recognition site, and a nonspecific sequence between the recognition and the cleaving sites (BciVI), were inhibited by PUVA treatment. Most of the other restriction enzymes used in this study, which do not have a 5'-TpA sequence at their restriction site, were not inhibited by PUVA treatment, although a 5'-TpA sequence is located adjacent (SmaI) or very close (BamHI, SacI and PstI) to the recognition and cleaving sites for these enzymes. Because SphI, which does not have 5'-TpA at its restriction site, was strongly inhibited by PUVA treatment, the 5'-CpA sequence is suggested to be a new binding site of psoralens after UVA irradiation.
Hanawa
F
F
Forestry and Forest Products Research Institute (FFPRI), P.O. Box 16, Tsukuba, Ibaraki 305-8687, Japan. fujinori@ffpri.affrc.go.jp
Okamoto
M
M
Towers
G H
GH
D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000037:antagonists & inhibitors*; D011564:Furocoumarins / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D011701:PUVA Therapy; D018462:Photobiology; D014466:Ultraviolet Rays
D011564:Furocoumarins; D004247:DNA; D004262:DNA Restriction Enzymes