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Originally published In Press as doi:10.1074/jbc.M507432200 on September 14, 2005
J. Biol. Chem., Vol. 280, Issue 46, 38457-38463, November 18, 2005
Higher Plant Plastids and Cyanobacteria Have Folate Carriers Related to Those of Trypanosomatids*
Sebastian M. J. Klaus ,
Edmund R. S. Kunji ,
Gale G. Bozzo ,
Alexandre Noiriel ,
Rocío Díaz de la Garza ,
Gilles J. C. Basset ,
Stéphane Ravanel¶,
Fabrice Rébeillé¶,
Jesse F. Gregory, III||, and
Andrew D. Hanson 1
From the
Horticultural Sciences and ||Food Science and Human Nutrition Departments, University of Florida, Gainesville, Florida 32611, the Medical Research Council, Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, United Kingdom, and the ¶Laboratoire de Physiologie Cellulaire Végétale, CNRS-Commissariat à l'Energie Atomique (Saclay, France) (CEA)-Institut National de la Recherche Agronomique-Université Joseph Fourier, CEA-Grenoble, F-38054 Grenoble Cedex 9, France
Cyanobacterial and plant genomes encode proteins with some similarity to the folate and biopterin transporters of the trypanosomatid parasite Leishmania. The Synechocystis slr0642 gene product and its closest Arabidopsis homolog, the At2g32040 gene product, are representative examples. Both have 12 probable transmembrane domains, and the At2g32040 protein has a predicted chloroplast transit peptide. When expressed in Escherichia coli pabA pabB or folE, mutants, which are unable to produce or take up folates, the slr0642 protein and a modified At2g32040 protein (truncated and fused to the N terminus of slr0642) enabled growth on 5-formyltetrahydrofolate or folic acid but not on 5-formyltetrahydrofolate triglutamate, demonstrating that both proteins mediate folate monoglutamate transport. Both proteins also mediate transport of the antifolate analogs methotrexate and aminopterin, as evidenced by their ability to greatly increase the sensitivity of E. coli to these inhibitors. The full-length At2g32040 polypeptide was translocated into isolated pea chloroplasts and, when fused to green fluorescent protein, directed the passenger protein to the envelope of Arabidopsis chloroplasts in transient expression experiments. At2g32040 transcripts were present at similar levels in roots and aerial organs, indicating that the protein occurs in non-green plastids as well as chloroplasts. Insertional inactivation of At2g32040 significantly raised the total folate content of chloroplasts and lowered the proportion of 5-methyltetrahydrofolate but did not discernibly affect growth. These findings establish conservation of function among folate and biopterin transporter family proteins from three kingdoms of life.
Received for publication, July 8, 2005
, and in revised form, August 19, 2005.
* This work was supported in part by the Florida Agricultural Experimental Station, by an endowment from the C. V. Griffin, Sr. Foundation, and by National Institutes of Health Grant R01 GM071382-01 and approved for publication as Journal Series no. R-10984. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. 1 and Table I.
1 To whom correspondence should be addressed: Horticultural Sciences Dept., University of Florida, Gainesville, FL 32611. Tel.: 352-392-1928 (ext. 334); Fax: 352-392-5653; E-mail: adha{at}mail.ifas.ufl.edu.

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