Drosophila mitochondrial transcription factor A: Characterization of its cDNA and expression pattern during development

We cloned a cDNA for Drosophila mitochondrial transcription factor A (D-mtT FA) and characterized the recombinant protein. In Drosophila Kc cells, D-mt TFA was localized in the mitochondria, but not in the nucleus. By repetitiv e precipitation with His-tag and PCR amplification, the consensus nucleotid e sequence for D-mtTFA-binding was determined to be 5 ' -TTATC/G. The bindi ng sequence was found to be clustered in the A + T region of mitochondrial DNA which is suggested to be a replication origin and promoter region for l ight strand and heavy strand. We found a DNA replication-related element (D RE)-like sequence located upstream of the transcription initiation site of the D-mtTFA gene and obtained results indicating that DRE-binding factor (D REF) can bind to the DRE-like sequence of the D-mtTFA gene. The data sugges t that transcription of the D-mtTFA gene is under control of the DRE/DREF r egulatory system. Based on these results, the functions of D-mtTFA were dis cussed in relation to mitochondrial biogenesis of Drosophila melanogaster. (C) 2001 Academic Press.