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Nucleic Acids Research, 2003, Vol. 31, No. 6 1597-1604
© 2003 Oxford University Press

DmTTF, a novel mitochondrial transcription termination factor that recognises two sequences of Drosophila melanogaster mitochondrial DNA

Marina Roberti1, Paola Loguercio Polosa1, Francesco Bruni1, Clara Musicco2, Maria Nicola Gadaleta1,2 and Palmiro Cantatore1,2

1 Dipartimento di Biochimica e Biologia Molecolare, Università di Bari and 2 Istituto di Biomembrane e Bioenergetica, CNR, Via Orabona 4, 70125 Bari, Italy

*To whom correspondence should be addressed at Dipartimento di Biochimica e Biologia Molecolare, Università di Bari, Via Orabona 4, 70125 Bari, Italy. Tel: +39 080 5443378; Fax: +39 080 5443403; Email: p.cantatore{at}biologia.uniba.it

Using a combination of bioinformatic and molecular biology approaches a Drosophila melanogaster protein, DmTTF, has been identified, which exhibits sequence and structural similarity with two mitochondrial transcription termination factors, mTERF (human) and mtDBP (sea urchin). Import/processing assays indicate that DmTTF is synthesised as a precursor of 410 amino acids and is imported into mitochondria, giving rise to a mature product of 366 residues. Band-shift and DNase I protection experiments show that DmTTF binds two homologous, short, non-coding sequences of Drosophila mitochondrial DNA, located at the 3' end of blocks of genes transcribed on opposite strands. The location of the target sequences coincides with that of two of the putative transcription termination sites previously hypothesised. These results indicate that DmTTF is the termination factor of mitochondrial transcription in Drosophila. The existence of two DmTTF binding sites might serve not only to stop transcription but also to control the overlapping of a large number of transcripts generated by the peculiar transcription mechanism operating in this organism.


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