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The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription

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 interferenc...

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Autores principales: Roberti, Marina, Bruni, Francesco, Polosa, Paola Loguercio, Gadaleta, Maria Nicola, Cantatore, Palmiro
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1450328/
https://www.ncbi.nlm.nih.gov/pubmed/16648357
http://dx.doi.org/10.1093/nar/gkl181
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author Roberti, Marina
Bruni, Francesco
Polosa, Paola Loguercio
Gadaleta, Maria Nicola
Cantatore, Palmiro
author_facet Roberti, Marina
Bruni, Francesco
Polosa, Paola Loguercio
Gadaleta, Maria Nicola
Cantatore, Palmiro
author_sort Roberti, Marina
collection PubMed
description 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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spelling pubmed-14503282006-05-03 The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription Roberti, Marina Bruni, Francesco Polosa, Paola Loguercio Gadaleta, Maria Nicola Cantatore, Palmiro Nucleic Acids Res Article 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. Oxford University Press 2006 2006-04-28 /pmc/articles/PMC1450328/ /pubmed/16648357 http://dx.doi.org/10.1093/nar/gkl181 Text en © The Author 2006. Published by Oxford University Press. All rights reserved
spellingShingle Article
Roberti, Marina
Bruni, Francesco
Polosa, Paola Loguercio
Gadaleta, Maria Nicola
Cantatore, Palmiro
The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription
title The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription
title_full The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription
title_fullStr The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription
title_full_unstemmed The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription
title_short The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription
title_sort drosophila termination factor dmttf regulates in vivo mitochondrial transcription
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1450328/
https://www.ncbi.nlm.nih.gov/pubmed/16648357
http://dx.doi.org/10.1093/nar/gkl181
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