Elongation by Drosophila RNA polymerase II. Transcription of 3'-extended DNA templates

Ann E Sluder; David H Price; Arno L Greenleaf

doi:10.1016/S0021-9258(19)81605-8

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Elongation by Drosophila RNA polymerase II. Transcription of 3'-extended DNA templates

Journal article

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Elongation by Drosophila RNA polymerase II. Transcription of 3'-extended DNA templates

Ann E Sluder, David H Price and Arno L Greenleaf

The Journal of biological chemistry, Vol.263(20), pp.9917-9925

07/15/1988

DOI: 10.1016/S0021-9258(19)81605-8

PMID: 2454924

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Abstract

RNA polymerase II will efficiently initiate transcription on linear duplex DNA which has been extended at its 3' ends by the addition of short stretches of polydeoxycytidine (Kadesch, T. R., and Chamberlin, M. J. (1982) J. Biol. Chem. 257, 5286-5295). We have used such dC-tailed templates to identify factors affecting elongation by Drosophila RNA polymerase II (Price, D. H., Sluder, A. E., and Greenleaf, A. L. (1987) J. Biol. Chem. 262, 3244-3255). While studying these factors we have observed two unexpected characteristics of transcription of the tailed templates. First, we found that RNA polymerase II encountered a strong pause site after the incorporation of 14 nucleotides. This pausing was observed on all templates examined and with RNA polymerase II from a variety of sources. In addition, we found that ammonium ions markedly stimulated the polymerase, increasing both the efficiency with which the enzyme left the 14 base pause site and the subsequent rate of elongation. A factor previously shown to affect transcription of dC-tailed templates (factor 4, Price, D. H., Sluder, A. E., and Greenleaf, A. L. (1987) J. Biol. Chem. 262, 3244-3255) was found to cause transcript displacement and to stimulate the elongation rate approximately 2-fold. This factor copurified with an RNase H activity, and a model is presented for the mechanism of transcript displacement by RNase H. The observations presented here form a basis for further analysis of RNA polymerase II elongation and its modulation by transcription factors. They should also aid in the interpretation of other experiments in which dC-tailed templates are used.

Ammonium Sulfate - pharmacology

Transcription, Genetic - drug effects

Endoribonucleases - metabolism

Templates, Genetic

Ribonuclease H

DNA - metabolism

RNA Polymerase II - metabolism

Nucleotides - metabolism

Nucleic Acid Heteroduplexes - metabolism

Drosophila - enzymology

RNA - genetics

Animals

Deoxycytidine - metabolism

Ribonuclease, Pancreatic - metabolism

Potassium Chloride - pharmacology

Kinetics

RNA - metabolism

Details

Title: Subtitle: Elongation by Drosophila RNA polymerase II. Transcription of 3'-extended DNA templates
Creators: Ann E Sluder - Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710
David H Price
Arno L Greenleaf
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.263(20), pp.9917-9925
DOI: 10.1016/S0021-9258(19)81605-8
PMID: 2454924
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: GM35500 / NIGMS NIH HHS GM28078 / NIGMS NIH HHS
Language: English
Date published: 07/15/1988
Academic Unit: Biochemistry and Molecular Biology
Record Identifier: 9984025252302771

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