Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published June 15, 1987 | Published
Journal Article Open

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

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.

Additional Information

© 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.

Attached Files

Published - NARpnas87.pdf

Files

NARpnas87.pdf
Files (1.5 MB)
Name Size Download all
md5:2503603e741d5ed602427cdcc2d80a82
1.5 MB Preview Download

Additional details

Created:
August 22, 2023
Modified:
October 13, 2023