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Nucleic Acids Research, 2003, Vol. 31, No. 17 5039-5047
© 2003 Oxford University Press

Import of mitochondrial transcription factor A (TFAM) into rat liver mitochondria stimulates transcription of mitochondrial DNA

Heike L. Garstka1, Wolfgang E. Schmitt1, Jeanette Schultz1, Bettina Sogl1, Barbara Silakowski2, Acisclo Pérez-Martos3, Julio Montoya3 and Rudolf J. Wiesner*,1,4

1 Department of Physiology II and 2 Zentrum für Molekulare Biologie (ZMBH), University of Heidelberg, Im Neuenheimer Feld, D-69120 Heidelberg, Germany, 3 Department of Biochemistry and Molecular and Cellular Biology, University of Zaragoza, Miguel Servet 177, E-50013 Zaragoza, Spain and 4 Institute of Vegetative Physiology, Medical Faculty, University of Köln, Robert-Koch-Strasse 39, D-50931 Köln, Germany

*To whom correspondence should be addressed at Institute of Vegetative Physiology, Medical Faculty, University of Köln, Robert-Koch-Strasse 39, D-50931 Köln, Germany. Tel: +49 221 478 3610; Fax: +49 221 478 3538; Email: rudolf.wiesner{at}uni-koeln.de
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

Mitochondrial transcription factor A (TFAM) has been shown to stimulate transcription from mitochondrial DNA promoters in vitro. In order to determine whether changes in TFAM levels also regulate RNA synthesis in situ, recombinant human precursor proteins were imported into the matrix of rat liver mitochondria. After uptake of wt-TFAM, incorporation of [{alpha}-32P]UTP into mitochondrial mRNAs as well as rRNAs was increased 2-fold (P < 0.05), whereas import of truncated TFAM lacking 25 amino acids at the C-terminus had no effect. Import of wt-TFAM into liver mitochondria from hypothyroid rats stimulated RNA synthesis up to 4-fold. We conclude that the rate of transcription is submaximal in freshly isolated rat liver mitochondria and that increasing intra-mitochondrial TFAM levels is sufficient for stimulation. The low transcription rate associated with the hypothyroid state observed in vivo as well as in organello seems to be a result of low TFAM levels, which can be recovered by treating animals with T3 in vivo or by importing TFAM in organello. Thus, this protein meets the criteria for being a key factor in regulating mitochondrial gene expression in vivo.


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