A heteroduplex template circumvents the energetic requirement for ATP during activated transcription by RNA polymerase II.
Unlike most eukaryotic and prokaryotic RNA polymerases, promoter-specific transcription by RNA polymerase II requires hydrolysis of the ATP beta-gamma phosphoanhydride bond. Here we show that a template containing a 10-base pair DNA mismatch encompassing the start site circumvents this requirement such that the non-hydrolyzable ATP analogues, ATP gamma S (adenosine 5'-O-(thiotriphosphate)) and AMP-PNP (adenyl-5'-yl imidodiphosphate), support both basal and GAL4-VP16-activated transcription in a reconstituted HeLa cell in vitro transcription system. The results imply that ATP regulates either opening of the template at the start site or a closely associated step.
Tantin
D
D
Molecular Biology Institute, UCLA School of Medicine 90024.
Carey
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes; D012319:RNA Polymerase II / Q000378:metabolism*; D013698:Templates, Genetic; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D017931:DNA Primers; D009692:Nucleic Acid Heteroduplexes; D014157:Transcription Factors; D000255:Adenosine Triphosphate; D004247:DNA; D012319:RNA Polymerase II