The Drosophila termination factor DmTTF regulates in vivo mitochondrial transcription.

Nucleic Acids Research
M RobertiP Cantatore

Abstract

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 addi...Continue Reading

References

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Apr 23, 2005·Biochemical and Biophysical Research Communications·Marina RobertiPalmiro Cantatore
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Citations

Mar 1, 2012·Journal of Molecular Evolution·Juliette ArabiAlexandre Hassanin
Jun 15, 2010·Nature Structural & Molecular Biology·Nereida Jiménez-MenéndezMaria Solà
Mar 30, 2007·Nucleic Acids Research·Paola Loguercio PolosaPalmiro Cantatore
Jan 11, 2013·Nucleic Acids Research·Yulia V Surovtseva, Gerald S Shadel
Sep 22, 2010·PloS One·Shu-Jun WeiJun-Hua He
Jul 28, 2011·The Plant Journal : for Cell and Molecular Biology·Víctor QuesadaJosé Luis Micol
Jun 15, 2010·Environmental and Molecular Mutagenesis·Timothy E Shutt, Gerald S Shadel
Dec 6, 2011·Biochimica Et Biophysica Acta·Kip E Guja, Miguel Garcia-Diaz
Apr 16, 2009·Biochimica Et Biophysica Acta·Marina RobertiPalmiro Cantatore
Aug 1, 2010·Biomolecular Concepts·Marina RobertiPalmiro Cantatore
Jul 13, 2012·Mitochondrion·Francesco BruniMarina Roberti
Jun 24, 2014·Molecular Systems Biology·Atsushi FukuohHoward T Jacobs
Jun 26, 2010·Molecular Biology Reports·Anne K HyvärinenHoward T Jacobs

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Methods Mentioned

BETA
PCR
electrophoresis
in vitro transcription

Software Mentioned

ImageQuant
Dissociation Curves
Primer Express

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