First published online July 18, 2002; 10.1104/pp.003798
Plant Physiol, August 2002, Vol. 129, pp. 1581-1591
Expression and Molecular Analysis of the Arabidopsis
DXR Gene Encoding 1-Deoxy-D-Xylulose
5-Phosphate Reductoisomerase, the First Committed Enzyme of the
2-C-Methyl-D-Erythritol 4-Phosphate
Pathway1
Lorenzo
Carretero-Paulet,
Iván
Ahumada,2
Nuria
Cunillera,
Manuel
Rodríguez-Concepción,
Albert
Ferrer,
Albert
Boronat, and
Narciso
Campos*
Departament de Bioquímica i Biologia Molecular, Facultat de
Química, Universitat de Barcelona, C/Martí i
Franquès 1, 08028 Barcelona, Spain (L.C.-P., I.A., M.R.-C., A.B.,
Na.C.); and Departament de Bioquímica i Biologia Molecular,
Facultat de Farmàcia, Universitat de Barcelona, Avinguda
Diagonal 643, 08028 Barcelona, Spain (Nu.C., A.F.)
1-Deoxy-D-xylulose 5-phosphate reductoisomerase
(DXR) catalyzes the first committed step of the
2-C-methyl-D-erythritol 4-phosphate pathway
for isoprenoid biosynthesis. In Arabidopsis, DXR is encoded by a
single-copy gene. We have cloned a full-length cDNA corresponding to
this gene. A comparative analysis of all plant DXR sequences known to
date predicted an N-terminal transit peptide for plastids, with a
conserved cleavage site, and a conserved proline-rich region at the N
terminus of the mature protein, which is not present in the prokaryotic
DXR homologs. We demonstrate that Arabidopsis DXR is targeted to
plastids and localizes into chloroplasts of leaf cells. The presence of
the proline-rich region in the mature Arabidopsis DXR was confirmed by
detection with a specific antibody. A proof of the enzymatic function
of this protein was obtained by complementation of an
Escherichia coli mutant defective in DXR activity. The
expression pattern of  -glucuronidase, driven by the
DXR promoter in Arabidopsis transgenic plants, together with the tissue distribution of DXR transcript and
protein, revealed developmental and environmental regulation of the
DXR gene. The expression pattern of the
DXR gene parallels that of the Arabidopsis 1-deoxy-D-xylulose 5-phosphate synthase gene, but the
former is slightly more restricted. These genes are expressed in most
organs of the plant including roots, with higher levels in seedlings and inflorescences. The block of the
2-C-methyl-D-erythritol 4-phosphate pathway
in Arabidopsis seedlings with fosmidomycin led to a rapid accumulation
of DXR protein, whereas the 1-deoxy-D-xylulose 5-phosphate synthase protein level was not altered. Our results are consistent with
the participation of the Arabidopsis DXR gene in the control of the
2-C-methyl-D-erythritol 4-phosphate pathway.
1
This work was supported by the Spanish
Ministerio de Educación y Cultura (grant no. BIO2000-0334), by
the Generalitat de Catalunya (grant no. CIRIT 1999SGR-00032), and by
the Agencia Española de Cooperación Internacional
(predoctoral MUTIS fellowship to I.A.).
2
Present address: Instituto de Biología Vegetal y
Biotecnología, Universidad de Talca, 2 Norte 685, Casilla 747, Talca, Chile.
*
Corresponding author; e-mail campos{at}sun.bq.ub.es; fax
34-93-4021219.
© 2002 American Society of Plant Physiologists
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