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J Biol Chem, Vol. 275, Issue 17, 13134-13141, April 28, 2000

Sequential Serum-dependent Activation of CREB and NRF-1 Leads to Enhanced Mitochondrial Respiration through the Induction of Cytochrome c*

Ronald P. HerzigDagger , Salvatore Scacco§, and Richard C. ScarpullaDagger

From the Dagger  Department of Cell and Molecular Biology, Northwestern Medical School, Chicago, Illinois 60611 and the § Institute of Medical Biochemistry and Chemistry, University of Bari, Piazza G. Cesare, 70124 Bari, Italy

Progression through the cell cycle requires ATP for protein synthesis, cytoskeletal rearrangement, chromatin remodeling, and protein degradation. The mechanisms by which mammalian cells increase respiratory capacity and ATP production in preparation for cell division are largely unexplored. Here, we demonstrate that serum induction of cytochrome c mRNA and processed protein in quiescent BALB/3T3 fibroblasts is associated with a marked increase in mitochondrial respiration. Cytochrome c was induced in the absence of any increase in citrate synthase activity or in subunit IV of the cytochrome c oxidase complex mRNA or protein, indicating that the enhanced respiratory rate did not require a general increase in mitochondrial biogenesis or respiratory chain expression. Transfections with a series of cytochrome c promoter mutants showed that both nuclear respiratory factor 1 (NRF-1) and cAMP-response element-binding protein (CREB) binding sites contributed equally to induced expression by serum. Moreover, CREB and NRF-1 were phosphorylated sequentially in response to serum, and the NRF-1 phosphorylation was accompanied by an increase in its ability to trans-activate target gene expression. The results demonstrate that the differential transcriptional expression of cytochrome c, through sequential transcription factor phosphorylations, leads to enhanced mitochondrial respiratory capacity upon serum-induced entry to the cell cycle.


* This work was supported by United States Public Health Service Grant GM32525-17 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Cell and Molecular Biology, Northwestern Medical School, 303 East Chicago Ave., Chicago, IL 60611. Tel.: (312) 503-2946; Fax: (312) 503-0798; E-mail: rsc248@nwu.edu.


Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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