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First published online June 26, 2003; 10.1104/pp.103.023242

Plant Physiology 132:1678-1687 (2003)
© 2003 American Society of Plant Biologists

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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Mitochondrial Phosphatidylserine Decarboxylase from Higher Plants. Functional Complementation in Yeast, Localization in Plants, and Overexpression in Arabidopsis1

Denis Rontein2, Wen-I Wu2, Dennis R. Voelker and Andrew D. Hanson*

Horticultural Sciences Department, University of Florida, Gainesville, Florida 32611 (D.R., A.D.H.); and Program in Cell Biology, Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206 (W.-I.W., D.R.V.)

Plants are known to synthesize ethanolamine (Etn) moieties by decarboxylation of free serine (Ser), but there is also some evidence for phosphatidyl-Ser (Ptd-Ser) decarboxylation. Database searches identified diverse plant cDNAs and an Arabidopsis gene encoding 50-kD proteins homologous to yeast (Saccharomyces cerevisiae) and mammalian mitochondrial Ptd-Ser decarboxylases (PSDs). Like the latter, the plant proteins have putative mitochondrial targeting and inner membrane sorting sequences and contain near the C terminus a Glycine-Serine-Threonine motif corresponding to the site of proteolysis and catalytic pyruvoyl residue formation. A truncated tomato (Lycopersicon esculentum) cDNA lacking the targeting sequence and a chimeric construct in which the targeting and sorting sequences were replaced by those from yeast PSD1 both complemented the Etn requirement of a yeast psd1 psd2 mutant, and PSD activity was detected in the mitochondria of the complemented cells. Immunoblot analysis of potato (Solanum tuberosum) mitochondria demonstrated that PSD is located in mitochondrial membranes, and mRNA analysis in Arabidopsis showed that the mitochondrial PSD gene is expressed at low levels throughout the plant. An Arabidopsis knockup mutant grew normally but had 6- to 13-fold more mitochondrial PSD mRNA and 9-fold more mitochondrial PSD activity. Total membrane PSD activity was, however, unchanged in the mutant, showing mitochondrial activity to be a minor part of the total. These results establish that plants can synthesize Etn moieties via a phospholipid pathway and have both mitochondrial and extramitochondrial PSDs. They also indicate that mitochondrial PSD is an important housekeeping enzyme whose expression is strongly regulated at the transcriptional level.


Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.103.023242.

1 This work was supported in part by the Florida Agricultural Experiment Station, by an endowment from the C.V. Griffin, Sr. Foundation, by the U.S. Department of Agriculture National Research Initiative Competitive Grants Program (grant no. 2001–35100–10620 to A.D.H.), and by the National Institutes of Health (grant no. GM 32453 to D.R.V.), and has been approved for publication as journal series no. R–09358.

2 These authors contributed equally to the paper.

* Corresponding author; e-mail adha{at}mail.ifas.ufl.edu; fax 352–392–5653.

Received March 8, 2003; returned for revision April 2, 2003; accepted April 4, 2003.




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