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Unexpected sequences and structures of mtDNA required for efficient transcription from the first heavy-strand promoter

Akira Uchida, Divakaran Murugesapillai, Yao Wang, Maria F. Lodeiro, Shaan Prabhakar, Jamie J. Arnold, L. James Maher III, Mark C. Williams, View ORCID ProfileCraig E. Cameron
doi: https://doi.org/10.1101/129908
Akira Uchida
1 The Pennsylvania State University, Department of Biochemistry and Molecular Biology, University Park, PA 16802
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Divakaran Murugesapillai
2 Northeastern University, Department of Physics, Boston, MA 02115
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Yao Wang
1 The Pennsylvania State University, Department of Biochemistry and Molecular Biology, University Park, PA 16802
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Maria F. Lodeiro
1 The Pennsylvania State University, Department of Biochemistry and Molecular Biology, University Park, PA 16802
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Shaan Prabhakar
1 The Pennsylvania State University, Department of Biochemistry and Molecular Biology, University Park, PA 16802
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Jamie J. Arnold
1 The Pennsylvania State University, Department of Biochemistry and Molecular Biology, University Park, PA 16802
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L. James Maher III
3 Mayo Clinic College of Medicine, Department of Biochemistry and Molecular Biology, Rochester, MN 55905
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Mark C. Williams
2 Northeastern University, Department of Physics, Boston, MA 02115
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Craig E. Cameron
1 The Pennsylvania State University, Department of Biochemistry and Molecular Biology, University Park, PA 16802
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  • ORCID record for Craig E. Cameron
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Abstract

Human mtDNA contains three promoters, suggesting a need for differential expression of the mitochondrial genome. Studies of mitochondrial transcription have used a reductionist approach, perhaps masking differential regulation. Here we evaluate transcription from light-strand (LSP) and heavy-strand (HSP1) promoters using templates that mimic their natural context. These studies reveal sequences upstream, hypervariable in the human population (HVR3), and downstream of the HSP1 transcription start site required for maximal yield. The carboxy-terminal tail of TFAM is essential for activation of HSP1 but not LSP. Images of the template obtained by atomic force microscopy show that TFAM creates loops in a discrete region, the formation of which correlates with activation of HSP1; looping is lost in tail-deleted TFAM. Identification of HVR3 as a transcriptional regulatory element may contribute to between-individual variability in mitochondrial gene expression. The unique requirement of HSP1 for the TFAM tail may enable its regulation by post-translational modifications.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted April 24, 2017.
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Unexpected sequences and structures of mtDNA required for efficient transcription from the first heavy-strand promoter
Akira Uchida, Divakaran Murugesapillai, Yao Wang, Maria F. Lodeiro, Shaan Prabhakar, Jamie J. Arnold, L. James Maher III, Mark C. Williams, Craig E. Cameron
bioRxiv 129908; doi: https://doi.org/10.1101/129908
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Unexpected sequences and structures of mtDNA required for efficient transcription from the first heavy-strand promoter
Akira Uchida, Divakaran Murugesapillai, Yao Wang, Maria F. Lodeiro, Shaan Prabhakar, Jamie J. Arnold, L. James Maher III, Mark C. Williams, Craig E. Cameron
bioRxiv 129908; doi: https://doi.org/10.1101/129908

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