Abstract
The synthesis and enzymatic incorporation into RNA of the hydrogen bond degenerate nucleoside analogue 6-(beta-d-ribofuranosyl)-3, 4-dihydro-8H-pyrimido[4,5-c]-[1,2]oxazin-7-one (P) is described. The 5'-triphosphate of this analogue is readily incorporated by T3, T7 and SP6 RNA polymerases into RNA transcripts, being best incorporated in place of UTP, but also in place of CTP. When all the uridine residues in an HIV-1 TAR RNA transcript are replaced by P the transcript has similar characteristics to the wild-type TAR RNA, as demonstrated by similar melting temperatures and CD spectra. The P-substituted TAR transcript binds to the Tat peptide ADP-1 with only 4-fold lowered efficiency compared with wild-type TAR.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Circular Dichroism
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Cytidine Triphosphate / analogs & derivatives
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Cytidine Triphosphate / chemical synthesis
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Cytidine Triphosphate / metabolism*
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DNA-Directed RNA Polymerases / metabolism*
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HIV Long Terminal Repeat / genetics
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HIV-1 / genetics
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Molecular Sequence Data
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Nucleic Acid Denaturation
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RNA, Messenger / biosynthesis*
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RNA, Messenger / chemistry
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RNA, Viral / biosynthesis*
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RNA, Viral / chemistry
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Uridine Triphosphate / analogs & derivatives
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Uridine Triphosphate / chemical synthesis
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Uridine Triphosphate / metabolism*
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Viral Proteins
Substances
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RNA, Messenger
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RNA, Viral
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Viral Proteins
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Cytidine Triphosphate
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RNA polymerase SP6
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bacteriophage T3 RNA polymerase
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bacteriophage T7 RNA polymerase
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DNA-Directed RNA Polymerases
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Uridine Triphosphate