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Overexpression of MTERFD1 or MTERFD3 impairs the completion of mitochondrial DNA replication

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Abstract

The physiological roles of the mitochondrial transcription termination factor (mTERF) family are poorly understood. MTERF and its homologues influence transcriptional readthrough in vitro, but the extent to which they regulate mitochondrial RNA levels in vivo is unclear. In addition, MTERF was previously shown to promote replication pausing. To test their roles in mtDNA metabolism, we created cell-lines inducibly expressing epitope-tagged versions of two members of the mTERF family, MTERFD1 and MTERFD3, as well as shRNA constructs targeted at each. We confirmed mitochondrial targeting and lack of sequence-specific DNA binding for both factors. Over-expression of epitope-tagged MTERFD1 or MTERFD3 resulted in modest mtDNA copy-number depletion and an accumulation of specific mtDNA replication intermediates indicating an impairment of the terminal steps of replication. These findings further implicate the mTERF family in restraining replication fork progression and support the idea that they facilitate the orderly passage of replication and transcription machineries, thus contributing to genome stability.

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Acknowledgments

We thank Academy of Finland, Tampere University Hospital Medical Research Fund, the UK Medical Research Council, Sigrid Juselius Foundation and the EU for financial support, Aurelio Reyes, Steffi Goffart and Hans Spelbrink for useful discussions and Outi Kurronen, Merja Jokela and Tea Tuomela for technical assistance.

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Authors and Affiliations

  1. Institute of Medical Technology and Centre for Laboratory Medicine, Tampere University Hospital, 33014, University of Tampere, Finland

    Anne K. Hyvärinen, Jaakko L. O. Pohjoismäki & Howard T. Jacobs

  2. MRC Mitochondrial Biology Unit, Wellcome Trust-MRC Building, Hills Road, Cambridge, CB2 0XY, UK

    Ian J. Holt

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  1. Anne K. Hyvärinen
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  2. Jaakko L. O. Pohjoismäki
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  3. Ian J. Holt
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  4. Howard T. Jacobs
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Correspondence to Howard T. Jacobs.

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Hyvärinen, A.K., Pohjoismäki, J.L.O., Holt, I.J. et al. Overexpression of MTERFD1 or MTERFD3 impairs the completion of mitochondrial DNA replication. Mol Biol Rep 38, 1321–1328 (2011). https://doi.org/10.1007/s11033-010-0233-9

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  • DOI: https://doi.org/10.1007/s11033-010-0233-9

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