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Nucleic Acids Research 2006 34(7):2109-2116; doi:10.1093/nar/gkl181
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Published online 3 May 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Article

The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription

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

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

*To whom correspondence should be addressed. Tel: +39 080 5443378; Fax: +39 080 5443403; Email: p.cantatore{at}biologia.uniba.it

Received February 20, 2006. Revised March 22, 2006. Accepted March 22, 2006.

DmTTF is a Drosophila mitochondrial DNA-binding protein, which recognizes two sequences placed at the boundary of clusters of genes transcribed in opposite directions. To obtain in vivo evidences on the role of DmTTF, we characterized a DmTTF knock-down phenotype obtained by means of RNA interference in D.Mel-2 cells. By a combination of RNase protection and real-time RT–PCR experiments we found that knock-down determines remarkable changes in mitochondrial transcription. In particular, protein depletion increases not only the level of (+) and (–)strand RNAs mapping immediately after of the two protein-binding site, but also that of transcripts located further downstream. Unexpectedly, depletion of the protein also causes the decrease in the content of those transcripts mapping upstream of the protein target sites, including the two rRNAs. The changes in transcript level do not depend on a variation in mitochondrial DNA (mtDNA) content, since mtDNA copy number is unaffected by DmTTF depletion. This work shows conclusively that DmTTF arrests in vivo the progression of the mitochondrial RNA polymerase; this is the first ever-obtained evidence for an in vivo role of an animal mitochondrial transcription termination factor. In addition, the reported data provide interesting insights into the involvement of DmTTF in transcription initiation in Drosophila mitochondria.


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