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  Cross-strand binding of TFAM to a single mtDNA molecule forms the mitochondrial nucleoid.

Kukat, C., Davies, M. K., Wurm, C. A., Spåhr, H., Bonekamp, A., Kühl, I., et al. (2015). Cross-strand binding of TFAM to a single mtDNA molecule forms the mitochondrial nucleoid. Proceedings of the National Academy of Sciences of the United States of America, 112(36), 11288-11293. doi:10.1073/pnas.1512131112.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-468C-6 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-1759-F
Genre: Journal Article

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 Creators:
Kukat, C., Author
Davies, M. K., Author
Wurm, C. A.1, Author              
Spåhr, H., Author
Bonekamp, A., Author
Kühl, I., Author
Joos, F., Author
Loguercio-Polosa, P., Author
Park, B., Author
Poss, V., Author
Falkenberg, M., Author
Jakobs, S.2, Author              
Kühlbrandt, W., Author
Larsson, N. G., Author
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              
2Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578566              

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Free keywords: nucleoids; mitochondria; cryo-ET; STED; nanoscopy
 Abstract: Mammalian mitochondrial DNA (mtDNA) is packaged by mitochondrial transcription factor A (TFAM) into mitochondrial nucleoids that are of key importance in controlling the transmission and expression of mtDNA. Nucleoid ultrastructure is poorly defined, and therefore we used a combination of biochemistry, superresolution microscopy, and electron microscopy to show that mitochondrial nucleoids have an irregular ellipsoidal shape and typically contain a single copy of mtDNA. Rotary shadowing electron microscopy revealed that nucleoid formation in vitro is a multistep process initiated by TFAM aggregation and cross-strand binding. Superresolution microscopy of cultivated cells showed that increased mtDNA copy number increases nucleoid numbers without altering their sizes. Electron cryo-tomography visualized nucleoids at high resolution in isolated mammalian mitochondria and confirmed the sizes observed by superresolution microscopy of cell lines. We conclude that the fundamental organizational unit of the mitochondrial nucleoid is a single copy of mtDNA compacted by TFAM, and we suggest a packaging mechanism.

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Language(s): eng - English
 Dates: 2015-08-242015-09-08
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1073/pnas.1512131112
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Title: Proceedings of the National Academy of Sciences of the United States of America
Source Genre: Journal
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Pages: - Volume / Issue: 112 (36) Sequence Number: - Start / End Page: 11288 - 11293 Identifier: -