Abortive initiation by RNA polymerase II in vitro at the adenovirus 2 major late promoter.
J Biol Chem
; 262(31): 14990-7, 1987 Nov 05.
Article
en En
| MEDLINE
| ID: mdl-3667620
ABSTRACT
We have investigated the formation of the first phosphodiester bond by RNA polymerase II in vitro. The template was a cloned DNA bearing the adenovirus 2 major late promoter; transcription factors and RNA polymerase II were provided by a HeLa cell nuclear extract. Dinucleotide primers and single nucleoside triphosphates were used as substrates. We found that accurate initiation does occur when only one phosphodiester bond can be formed; however, all of the resulting dinucleotide-primed trimers are abortively initiated. Synthesis of the trimers by RNA polymerase II requires ATP or dATP and is sensitive to low concentrations of alpha-amanitin. Treatments which abolish the ability of the preinitiation complex to synthesize long RNAs also eliminate the ability to abortively initiate. Abortive initiation proceeds for at least one-half h at 25 degrees C, at which point up to 4 mol of transcript/mol of template have been synthesized. The level of abortive initiation (per template molecule) is not significantly reduced by 0.025% Sarkosyl or by 10-fold dilution of the reaction, consistent with the initiation complex remaining intact during abortive initiation.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
ARN Polimerasa II
/
Adenovirus Humanos
/
Regiones Promotoras Genéticas
/
Genes Virales
Límite:
Humans
Idioma:
En
Revista:
J Biol Chem
Año:
1987
Tipo del documento:
Article