The synthesis and enzymatic polymerization of 5-thio- and 5-methylmercurithio-pyrimidine nucleotides.
The 5-thio and 5-methylmercurithio derivatives of UTP, dUTP and dCTP have been synthesized and tested as substrates for nucleic acid polymerases. The 5-thio-nucleotides were polymerized inefficiently by both RNA polymerase and DNA polymerase I of Escherichia coli. The 5-methylmercurithio derivatives of dUTP and dCTP were, however, utilized by DNA polymerase I, an enzyme insensitive to mercurial compounds, although they were potent inhibitors of all other polymerases tested. While polymers containing the 5-thio substituent possess structural abnormalities, most likely interstrand disulfide bridges, polymers containing 5-methylmercurithio groups appear normal. The latter polynucleotides are readily separated from non-sulfated polymers by chromatography on mercuriagarose.
D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004229:Dithiothreitol; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D008767:Methylmercury Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D011742:Pyrimidine Nucleotides / Q000138:chemical synthesis* / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D013438:Sulfhydryl Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D013698:Templates, Genetic
D008767:Methylmercury Compounds; D011742:Pyrimidine Nucleotides; D013438:Sulfhydryl Compounds; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase; D004229:Dithiothreitol