First published online January 9, 2003; 10.1104/pp.009985
Plant Physiol, February 2003, Vol. 131, pp. 443-453
Genomic and Proteomic Analysis of Mitochondrial Carrier Proteins
in Arabidopsis1
A. Harvey
Millar* and
Joshua L.
Heazlewood
Plant Molecular Biology Group, School of Biomedical and Chemical
Sciences, The University of Western Australia, Crawley 6009, Western
Australia, Australia
Plant mitochondria maintain metabolic communication with the
cytosol through a family of carrier proteins. In Arabidopsis, a subset
of 45 putative genes encoding members of this family have been
identified based on generalized mitochondrial carrier features. No gene
clusters are apparent and few of the predicted protein products have
mitochondrial targeting sequences recognized by bioinformatic
predictors. Only nine genes are currently represented by more than 10 expressed sequence tags at The Institute for Genomic Research. Analyses
of public microarray experiments reveal differential expression
profiles of the more highly expressed members of this gene family in
different plant organs and in response to plant hormone application and
environmental stresses. A comparison of this Arabidopsis carrier subset
(45) to the yeast gene family (35) reveals 10 orthologous groups
between the two species. Recent surveys of the Arabidopsis
mitochondrial proteome by two-dimensional gel separations have not
identified any of these carrier proteins, presumably because of their
hydrophobicity and basicity. Isolating integral membrane proteins from
Arabidopsis mitochondria, using one-dimensional electrophoresis for
protein separation and tandem mass spectrometry-based sequencing of
doubly charged peptides, we have unequivocally identified specific
carrier gene products located in mitochondria. This approach has
identified six of the nine carriers represented highly in expressed
sequence tag databases: adenine nucleotide translocator (At3g8580 and
At5g13490), dicarboxylate/tricarboxylate carrier (At5g19760), phosphate
carrier (At5g14040), uncoupling protein (At3g54110), and a carrier gene
of unknown function (At4g01100). Overall, the combined transcript and
protein expression data indicates that only a small subset of the
carrier family of genes provide the majority of carrier proteins of
Arabidopsis mitochondria.
1
This work was supported by the Australian
Research Council Discovery Program (grants to A.H.M., D. Day, and J. Whelan). A.H.M. was supported by an Australian Research Council Queen
Elizabeth II fellowship.
*
Corresponding author; e-mail hmillar{at}cyllene.uwa.edu.au; fax
61-8-9280-1148.
© 2003 American Society of Plant Biologists
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