ORGN 511 |
| Zhen Huang and Nicolas Carrasco. Department of Biochemistry and Chemistry, Brooklyn College, 2900 Bedford Ave., Brooklyn, NY 11210 |
| The derivatization of DNA and RNA with suitable heavy atoms is a challenging problem in the field of nucleic acid X-ray crystallography. One approach to address this problem is to incorporate nucleotides containing specific heavy atoms covalently into DNA or RNA. We report here some studies on the novel synthesis of nucleoside triphosphates harboring the selenium functionality at the a-phosphate (NTPaSes) and their enzymatic incorporation into DNA or RNA using primer extension or in vitro transcription, respectively. To accomplish the synthesis of the NTPaSes, we exploited a nucleoside phosphite intermediate that was oxidized with selenium. Complete characterization of the analogs was carried out by 1H-NMR, 31P-NMR, mass spectroscopy, and HPLC. To investigate the suitability of NTPaSes for the enzymatic synthesis of DNA and RNA, we studied the efficiency of incorporation of the HPLC-isolated diastereomers of uridine and thymidine a-selenotriphosphates at various Mg2+, EDTA, and NTPaSe/NTP concentrations. The level of incorporation was analyzed by comparative gel electrophoresis using radioactive 31P-adenosine triphosphate. The results demonstrated that UTPaSe was an inhibitor of RNA polymerization, while TTPaSe promoted the polymerization of DNA efficiently. |
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Bioorganic, Molecular Recognition, Asymmetric Reactions and Syntheses
11:00 AM-1:00 PM, Wednesday, September 10, 2003 Javits Convention Center -- Hall 1B/1C, Poster
Division of Organic Chemistry |