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Inhibition of Restriction Enzyme's DNA Sequence Recognition by PUVA Treatment † ¶
Author(s) -
Hanawa Fujinori,
Okamoto Mamoru,
Towers G. H. Neil
Publication year - 2001
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1562/0031-8655(2001)0740269ioreds2.0.co2
Subject(s) - restriction enzyme , recognition sequence , psti , bamhi , restriction site , dna , restriction map , enzyme , sequence (biology) , microbiology and biotechnology , biology , chemistry , biochemistry , nucleic acid sequence
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 ( Kpn I, Xba I, Pme I and Dra I), and the noncleaving site ( Pac I) in recognition sites, or have two 5′‐TpA sequences at the recognition site, and a nonspecific sequence between the recognition and the cleaving sites ( Bci VI), 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 ( Sma I) or very close ( Bam HI, Sac I and Pst I) to the recognition and cleaving sites for these enzymes. Because Sph I, 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.