Abstract
The unit of inheritance for mitochondrial DNA (mtDNA) is a complex nucleoprotein structure termed the nucleoid. The organization of the nucleoid as well as its role in mtDNA replication remain largely unknown. Here, we show in Saccharomyces cerevisiae that at least two populations of nucleoids exist within the same mitochondrion and can be distinguished by their association with a discrete proteinaceous structure that spans the outer and inner mitochondrial membranes. Surprisingly, this two membrane-spanning structure (TMS) persists and self-replicates in the absence of mtDNA. We tested whether TMS functions to direct the replication of mtDNA. By monitoring BrdU incorporation, we observed that actively replicating nucleoids are associated exclusively with TMS. Consistent with TMS's role in mtDNA replication, we found that Mip1, the mtDNA polymerase, is also a stable component of TMS. Taken together, our observations reveal the existence of an autonomous two membrane-spanning mitochondrial replisome as well as provide a mechanism for how mtDNA replication and inheritance may be physically linked.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Cells, Cultured
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DNA Polymerase I / genetics
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DNA Polymerase I / metabolism
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DNA Replication / genetics*
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DNA, Mitochondrial / genetics*
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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DNA-Directed DNA Polymerase / genetics
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DNA-Directed DNA Polymerase / metabolism
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Green Fluorescent Proteins
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Indoles
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Intracellular Membranes / metabolism*
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Intracellular Membranes / ultrastructure
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Mitochondria / genetics*
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Mitochondria / metabolism
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Mitochondria / ultrastructure
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Mitochondrial Proteins
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Nucleoproteins / genetics
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Nucleoproteins / metabolism*
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / ultrastructure
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Transcription Factors / genetics
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Transcription Factors / metabolism
Substances
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ABF2 protein, S cerevisiae
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DNA, Mitochondrial
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DNA-Binding Proteins
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Indoles
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Luminescent Proteins
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MGM101 protein, S cerevisiae
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Mitochondrial Proteins
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Nuclear Proteins
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Nucleoproteins
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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Green Fluorescent Proteins
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DAPI
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DNA Polymerase I
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DNA-Directed DNA Polymerase
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MIP1 protein, S cerevisiae