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The synthesis and enzymatic polymerization of nucleotides containing mercury: potential tools for nucleic acid sequencing and structural analysis.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Dale, Roderic M. K. Livingston, Debra Ward, David C. |
| Copyright Year | 1973 |
| Abstract | A simple acetoxymercuration reaction for introducing covalently bound mercury atoms into nucleotides is described. The 5-mercuriacetate derivatives of UTP, CTP, dUTP, and dCTP, as well as the 7-mercuriacetate derivative of 7-deazaATP, have been prepared by this procedure and tested as substrates for nucleic acid polymerases. These nucleotides, in the absence of added mercaptan, are not polymerized and in most instances are potent enzyme inhibitors. However, conversion of these mercuriacetates to mercurithio compounds in situ by the addition of one of various mercaptans, yields nucleoside triphosphates that are excellent substrates for all polymerases tested: Escherichia coli and T7 RNA polymerases, DNA polymerase I of E. coli, DNA polymerase of avian myeloblastosis virus, and calf-thymus terminal deoxynucleotidyl transferase. By varying the mercaptan used to promote syntheses it is possible to access certain structural limitations in the enzyme's nucleoside triphosphate binding site. These mercurinucleotides appear to have a diversity of potential applications: (1) as heavy-atom reagents for crystallographic and microscopic studies; (2) as affinity probes for enzymes sensitive to sulfhydryl modification; (3) as steric probes of substrate-binding sites on enzymes; and (4) as reagents for forming covalent protein-polynucleotide complexes. |
| Starting Page | 1 |
| Ending Page | 2 |
| Page Count | 2 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://www.pnas.org/content/70/8/2238.full.pdf |
| PubMed reference number | 4365367v1 |
| Volume Number | 70 |
| Issue Number | 8 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | 2'-deoxycytidine 5'-triphosphate Aves Avian Myeloblastosis Virus Binding Sites Cell Nucleus Covalent Interaction DNA Nucleotidylexotransferase DNA-Directed RNA Polymerase Nucleic Acids Nucleic acid sequencing Nucleosides Nucleotides Polynucleotides Purine-Nucleoside Phosphorylase Reagents Sulfhydryl Compounds Thymic Tissue Transferase Uridine Triphosphate deoxyuridine triphosphate negative regulation of DNA-directed DNA polymerase activity nucleoside triphosphate |
| Content Type | Text |
| Resource Type | Article |
