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Originally published In Press as doi:10.1074/mcp.M300035-MCP200 on September 26, 2003.
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Molecular & Cellular Proteomics 2:1205-1216, 2003.
© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Protein Components of Mitochondrial DNA Nucleoids in Higher Eukaryotes*

Daniel F. Bogenhagen{ddagger},§, Yousong Wang{ddagger}, Ellen L. Shen{ddagger},¶ and Ryuji Kobayashi||,**

From the {ddagger} Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY 11794, and || Cold Spring Harbor Laboratories, Cold Spring Harbor, NY 11724

Mitochondrial DNA (mtDNA) is not packaged in nucleosomal particles, but has been reported to associate with the mitochondrial inner membrane. Gentle lysis of Xenopus oocyte mitochondria with nonionic detergent liberates a nucleoprotein complex containing mtDNA associated with a previously characterized DNA binding partner, mitochondrial transcription factor A (mtTFA), as well as a series of inner membrane proteins identified by sequencing. More extensive detergent treatment stripped most of these proteins from the DNA, leaving a limited number of proteins in a nucleoid core. Sequencing of the major proteins retained in association with mtDNA revealed the expected mtDNA binding proteins, mtTFA and mitochondrial single-stranded DNA binding protein (mtSSB), as well as four proteins not previously reported to associate with mtDNA. These include adenine nucleotide translocator 1, the lipoyl-containing E2 subunits of pyruvate dehydrogenase and branched chain {alpha}-ketoacid dehydrogenase and prohibitin 2. The association of several of these proteins with mtTFA-containing mtDNA nucleoids was confirmed by immunoprecipitation.

§ To whom correspondence should be addressed: Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY 11794-8651. Tel.: 631-444-3068; Fax: 631-444-3218; E-mail: dan{at}pharm.sunysb.edu.


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