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Specific requirement for ATP at an early step of in vitro transcription of human mitochondrial DNA

Narasimhan, Nalini and Attardi, Giuseppe (1987) Specific requirement for ATP at an early step of in vitro transcription of human mitochondrial DNA. Proceedings of the National Academy of Sciences of the United States of America, 84 (12). pp. 4078-4082. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:NARpnas87

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Abstract

The ATP concentrations allowing transcription of both heavy-and light-strand of human mtDNA in a HeLa cell mitochrondrial lysate were found to cover a broad range, with a maximum around 2.5 mM, and with reproducible differences in the ATP response curves for the two transcription events. Direct measurements showed that nonspecific ATP degradation during the assay did not account for the high ATP requirement. 5'-Adenylyl imidodiphosphate (p[NH]ppA), an ATP analog with a nonhydrolyzable ß-γ bond, was unable to substitute for ATP in supporting mtDNA transcription but greatly stimulated this transcription in the presence of a low concentration of exogenous ATP. Evidence was obtained indicating that p[NH]ppA did not support an early event in mtDNA transcription (formation of preinitiation complex or initiation), whereas this analog could substitute effectively for ATP in the subsequent elongation steps. These results pointed to a specific requirement for ATP at an early step of the transcription process.


Item Type:Article
Additional Information:Copyright © 1987 by the National Academy of Sciences. Contributed by Giuseppe Attardi, March 9, 1987. We are greatly indebted to David Shuey for helpful discussions and to George Gaines for providing us with the pmt-H8 clone of human mtDNA. The technical assistance of Arger Drew is gratefully acknowledged. These investigations were supported by National Institutes of Health Grant GM-11726 to G.A. and American Cancer Society Fellowship PF-2610 to N.N. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:imidonudceotides; promoters; heavy strand; light strand; RNA polymerase
Record Number:CaltechAUTHORS:NARpnas87
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:NARpnas87
Alternative URL:http://www.pnas.org/cgi/content/abstract/84/12/4078
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:1487
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:23 Jan 2006
Last Modified:26 Dec 2012 08:44

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