Transcription by SP6 RNA polymerase exhibits an ATP dependence that is influenced by promoter topology

Taylor, D. R., Mathews, M. B. (April 1993) Transcription by SP6 RNA polymerase exhibits an ATP dependence that is influenced by promoter topology. Nucleic Acids Res, 21 (8). pp. 1927-33. ISSN 0305-1048 (Print)

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URL: http://www.ncbi.nlm.nih.gov/pubmed/8493106
DOI: 10.1093/nar/21.8.1927

Abstract

Transcription of linearized DNA templates by SP6 RNA polymerase requires a higher concentration of ATP than of the other three nucleotides. This requirement is not shared by T7 RNA polymerase. The ATP requirement is partially relieved when the SP6 template is supercoiled but not when it is relaxed circular DNA. The effect of supercoiling is eliminated by replacement of the A.T rich sequence downstream from the SP6 promoter with a G.C rich sequence. Examination of the reaction products indicates that the ATP dependence of transcription from a linear template is not due to an ATPase activity or to the premature termination of transcription at low ATP concentration. These data suggest that the initiation of transcription by SP6 RNA polymerase requires partial denaturation of the template in the promoter-proximal region, and that this requirement can be satisfied by negative supercoiling or by increasing the ATP concentration. ATP also reduces, but does not eliminate, the abortive transcription that leads to the production of short, prematurely terminated transcripts by SP6 polymerase from supercoiled templates.

Item Type: Paper
Uncontrolled Keywords: Adenosine Triphosphate/ metabolism Bacteriophage T7/enzymology DNA-Directed RNA Polymerases/ metabolism Nucleic Acid Conformation Promoter Regions (Genetics) Research Support, U.S. Gov't, P.H.S. Salmonella Phages/ enzymology Transcription, Genetic
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > RNA polymerase
CSHL Authors:
Communities: CSHL labs
Depositing User: Matt Covey
Date: 25 April 1993
Date Deposited: 08 Apr 2016 18:33
Last Modified: 08 Nov 2017 16:12
PMCID: PMC309434
Related URLs:
URI: http://repository.cshl.edu/id/eprint/32583

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