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Sci Rep. 2015 Oct 19;5:15292. doi: 10.1038/srep15292.

Human Mitochondrial DNA-Protein Complexes Attach to a Cholesterol-Rich Membrane Structure.

Author information

1
Nijmegen Centre for Mitochondrial Disorders, RadboudUMC, Nijmegen, The Netherlands.
2
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
3
Faculty of Science and Engineering, Biochemistry. Åbo Akademi, Turku, Finland.
4
MRC Mitochondrial Biology Unit, Cambridge, UK.
5
Institute for Cell and Molecular Biosciences, Newcastle University Institute for Ageing, University of Newcastle, Newcastle upon Tyne, NE4 5PL, UK.

Abstract

The helicase Twinkle is indispensable for mtDNA replication in nucleoids. Previously, we showed that Twinkle is tightly membrane-associated even in the absence of mtDNA, which suggests that Twinkle is part of a membrane-attached replication platform. Here we show that this platform is a cholesterol-rich membrane structure. We fractionated mitochondrial membrane preparations on flotation gradients and show that membrane-associated nucleoids accumulate at the top of the gradient. This fraction was shown to be highly enriched in cholesterol, a lipid that is otherwise low abundant in mitochondria. In contrast, more common mitochondrial lipids, and abundant inner-membrane associated proteins concentrated in the bottom-half of these gradients. Gene silencing of ATAD3, a protein with proposed functions related to nucleoid and mitochondrial cholesterol homeostasis, modified the distribution of cholesterol and nucleoids in the gradient in an identical fashion. Both cholesterol and ATAD3 were previously shown to be enriched in ER-mitochondrial junctions, and we detect nucleoid components in biochemical isolates of these structures. Our data suggest an uncommon membrane composition that accommodates platforms for replicating mtDNA, and reconcile apparently disparate functions of ATAD3. We suggest that mtDNA replication platforms are organized in connection with ER-mitochondrial junctions, facilitated by a specialized membrane architecture involving mitochondrial cholesterol.

PMID:
26478270
PMCID:
PMC4609938
DOI:
10.1038/srep15292
[Indexed for MEDLINE]
Free PMC Article

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