Transcription of 2'-deoxy-2'-fluoro-modified and phosphorothioate-modified RNA templates by HIV-1 reverse transcriptase.
RNA templates yield the corresponding DNA in the presence of human immunodeficiency virus reverse transcriptase (HIV-1 RT). The purpose of this study was to determine whether RNA that was modified with either 2'-deoxy-2'-fluoro analogs or with internucleotide phosphorothioate linkages could serve as templates for HIV-1 RT. Modified RNA that contained either 2'-deoxy 2'-fluoro pyrimidine nucleoside analogs or internucleotide phosphorothioate diester linkages 5'- to pyrimidine nucleosides were enzymatically synthesized and tested for template activity with recombinant HIV-1 RT. RNA that was modified with either 2'-deoxy-2'-fluorouridine or with internucleotide phosphorothioate linkages 5'- to pyrimidines yielded full length HIV-1 reverse transcription products, with complete fidelity in transcription. RNA that was modified with 2'-deoxy-2'-fluorocytidine, either alone or in combination with 2'-deoxy-2'-fluorouridine, did not function as templates for HIV-1 RT, under the conditions reported here. The ability of 2'-deoxy-2'-fluoro-modified and phosphorothioate-modified RNA to serve as template for the RNA-dependent DNA polymerase of HIV-1 RT has not hitherto been reported.
Karim
A S
AS
Department of Neurosurgery, University of Arkansas for Medical Sciences, Little Rock 72205, USA.
Weltman
J K
JK
D001483:Base Sequence; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D054303:HIV Reverse Transcriptase / Q000235:genetics*; D012313:RNA / Q000737:chemistry / Q000235:genetics*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D017931:DNA Primers; D012313:RNA; D054303:HIV Reverse Transcriptase