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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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: | https://repository.cshl.edu/id/eprint/32583 |
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