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Published August 26, 2013 | Supplemental Material + Published
Journal Article Open

Genome-Wide Analysis Reveals Coating of the Mitochondrial Genome by TFAM

Abstract

Mitochondria contain a 16.6 kb circular genome encoding 13 proteins as well as mitochondrial tRNAs and rRNAs. Copies of the genome are organized into nucleoids containing both DNA and proteins, including the machinery required for mtDNA replication and transcription. The transcription factor TFAM is critical for initiation of transcription and replication of the genome, and is also thought to perform a packaging function. Although specific binding sites required for initiation of transcription have been identified in the D-loop, little is known about the characteristics of TFAM binding in its nonspecific packaging state. In addition, it is unclear whether TFAM also plays a role in the regulation of nuclear gene expression. Here we investigate these questions by using ChIP-seq to directly localize TFAM binding to DNA in human cells. Our results demonstrate that TFAM uniformly coats the whole mitochondrial genome, with no evidence of robust TFAM binding to the nuclear genome. Our study represents the first high-resolution assessment of TFAM binding on a genome-wide scale in human cells.

Additional Information

© 2013 Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received June 27, 2013; Accepted August 5, 2013; Published August 26, 2013. Funding: This work was supported by NIH grants GM062967 (DCC), U54 HG004576 (BJW), U54 HG006998 (BJW), the Beckman Foundation, and the Donald Bren Endowment. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Igor Antoshechkin and the Millard and Muriel Jacobs Genetics and Genomics Laboratory for assistance with library building and high-throughput sequencing. We thank Henry Amrhein and Diane Trout for computational assistance, Susan Ou for monoclonal antibody production, and Elizabeth Nelson for help with antibody characterization. Author Contributions: Conceived and designed the experiments: YEW DCC. Performed the experiments: YEW. Analyzed the data: YEW GKM. Wrote the manuscript: YEW GKM BJW DCC.

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