title: Polymerase synthesis of DNAs bearing vinyl groups in the major groove and their cleavage by restriction endonucleases.
abstract: DNA molecules containing 5-vinyluracil, 5-vinylcytosine, or 7-deaza-7-vinyladenine were prepared by polymerase incorporation of the corresponding vinyl-modified 2'-deoxyribonucleoside triphosphates, and the influence of the vinyl group in the major groove of DNA on the cleavage by diverse type II restriction endonucleases (REs) was studied. The presence of 5-vinyluracil was tolerated by most of the REs, whereas only some REs were able to cleave sequences containing 7-deaza-7-vinyladenine. The enzyme ScaI was found to cleave DNA containing 5-vinylcytosine efficiently but not DNA containing the related 5-ethynylcytosine. All other REs failed to cleave sequences containing any cytosine modifications.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Science Czech Republic, Gilead Sciences and IOCB Research Center, Flemingovo nám. 2, 16610 Prague 6 (Czech Republic).
authors.forename: Michaela
authors.initials: M
authors.lastname: Mačková
authors.forename: Radek
authors.initials: R
authors.lastname: Pohl
authors.forename: Michal
authors.initials: M
authors.lastname: Hocek
mesh_terms: D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D053837:DNA Cleavage; D004262:DNA Restriction Enzymes / Q000378:metabolism*
chemical_list: D004247:DNA; D004262:DNA Restriction Enzymes
keywords: DNA; DNA polymerases; DNA-protein interactions; nucleotides; restriction endonucleases