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J. Biol. Chem., Vol. 269, Issue 1, 105-113, Jan, 1994
L Gopalakrishnan and RC Scarpulla
In vertebrate organisms, the molecular mechanisms by which extracellular
signals regulate mitochondrial function and biogenesis are largely unknown.
We have previously identified multiple cis-acting elements in both
cytochrome c and cytochrome oxidase subunit IV (COXIV) genes that are
likely targets for the regulated expression of respiratory chain
components. We now demonstrate that cytochrome c but not COXIV mRNA is
induced by cAMP through a mechanism involving transcriptional activation.
Maximal induction occurs within 3 h and does not require de novo protein
synthesis. The differential response of these genes is mediated by two
distinct cAMP response elements (CREs) in the cytochrome c promoter region.
Both elements function independently to drive cAMP-dependent expression
from a heterologous promoter and within the proper cytochrome c promoter
context. In addition, the binding properties of both elements to nuclear
factors were characterized by competition DNase I footprinting, methylation
interference footprinting, site-directed mutagenesis, and UV-induced
DNA-protein cross-linking. The results are all consistent with the specific
recognition of both CREs by CRE binding protein (CREB). A highly purified
preparation of recombinant CREB formed a specific complex with each of the
cytochrome c CREs identical to that formed with a crude nuclear fraction.
In addition, the trans-activation of cytochrome c gene expression by
recombinant CREB and protein kinase A in transfected cells was completely
dependent on functional CREs within the promoter. These results establish
that respiratory chain gene expression can be regulated directly by cAMP
through a CREB-dependent signal transduction pathway.
Differential regulation of respiratory chain subunits by a CREB- dependent signal transduction pathway. Role of cyclic AMP in cytochrome c and COXIV gene expression
Department of Cell, Molecular, and Structural Biology, Northwestern University Medical School, Chicago, Illinois 60611.
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