Issue 5, 2011

Shot-gun proteomic analysis of mitochondrial D-loop DNA binding proteins: identification of mitochondrial histones

Abstract

Transcription and replication of mitochondrial DNA (mtDNA) are regulated by nuclear DNA-encoded proteins that are targeted into mitochondria. A decrease in mtDNA copy number results in mitochondrial dysfunction, which may lead to insulin resistance and metabolic syndromes. We analyzed mitochondrial proteins that physically bind to human mitochondrial D-loop DNA using a shot-gun proteomics approach following protein enrichment by D-loop DNA-linked affinity chromatography. A total of 152 D-loop DNA binding proteins were identified by peptide sequencing using ultra high pressure capillary reverse-phase liquid chromatography/tandem mass spectrometry. Bioinformatic analysis showed that 68 were mitochondrial proteins, 96 were DNA/RNA/protein binding proteins and 114 proteins might form a complex viaprotein–protein interactions. Histone family members of H1, H2A, H2B, H3, and H4, were detected in abundance among them. In particular, histones H2A and H2B were present in the mitochondrial membrane as integral membrane proteins and not bound directly to mtDNA inside the organelle. Histones H1.2, H3 and H4 were associated with the outer mitochondrial membrane. Silencing of H2AX expression inhibited mitochondrial protein transport. Our data suggests that many mitochondrial proteins may reside in multiple subcellular compartments like H2AX and exert multiple functions.

Graphical abstract: Shot-gun proteomic analysis of mitochondrial D-loop DNA binding proteins: identification of mitochondrial histones

Supplementary files

Article information

Article type
Paper
Submitted
12 Nov 2010
Accepted
21 Jan 2011
First published
28 Feb 2011

Mol. BioSyst., 2011,7, 1523-1536

Shot-gun proteomic analysis of mitochondrial D-loop DNA binding proteins: identification of mitochondrial histones

Y. Choi, J. Hoon Jeong, H. Min, H. Jung, D. Hwang, S. Lee and Y. Kim Pak, Mol. BioSyst., 2011, 7, 1523 DOI: 10.1039/C0MB00277A

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