Abstract
Higher plant nitrate reductases (NRs) carry an N-terminal domain whose sequence is not conserved in NRs from other organisms. A gene composed of a full-length tobacco NR cDNA with an internal deletion of 168 bp in the 5' end fused to the cauliflower mosaic virus 35S promoter and appropriate termination signals was constructed and designated as delta NR. An NR-deficient mutant of Nicotiana plumbaginifolia was transformed with this delta NR gene. In transgenic plants expressing this construct, NR activity was restored and normal growth resulted. Apart from a higher thermosensitivity, no appreciable modification of the kinetic parameters of the enzyme was detectable. The post-transcriptional regulation of NR by light was abolished in delta NR transformants. Consequently, deregulated production of glutamine and asparagine was detected in delta NR transformants. The absence of in vitro delta NR activity modulation by ATP suggests the impairment of delta NR phosphorylation and thereby suppression of delta NR post-translational regulation. These data imply that post-transcriptional control of NR expression is important for the flow of the nitrate assimilatory pathway.
MeSH Terms
Adenosine Triphosphate/pharmacology
Amino Acid Sequence
Asparagine/biosynthesis
Base Sequence
Conserved Sequence
Darkness
Gene Expression Regulation, Enzymologic/radiation effects
Gene Expression Regulation, Plant/radiation effects
Genes, Plant/genetics
Genetic Complementation Test
Glutamine/biosynthesis
Homozygote
Light
Molecular Sequence Data
Nitrate Reductase
Nitrate Reductases/biosynthesis,drug effects,genetics
Peptides/analysis
Phosphorylation
Plants, Genetically Modified
Plants, Toxic
Protein Processing, Post-Translational
RNA Processing, Post-Transcriptional
RNA, Plant/analysis
Sequence Deletion
Sequence Homology, Amino Acid
Tobacco/enzymology,genetics
Yeasts/genetics
Chemicals
Peptides
RNA, Plant
Glutamine
Asparagine
Adenosine Triphosphate
Nitrate Reductases
Nitrate Reductase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Nussaume L
Laboratoire de Biologie Cellulaire, Institut National de la Recherche Agronomique, Centre de Versailles, Versailles, France.
Vincentz M
Meyer C
Boutin J P
Caboche M
References (16)
16 references, click to expand
-
Rapid Modulation of Spinach Leaf Nitrate Reductase Activity by Photosynthesis : I. Modulation in Vivo by CO(2) Availability.
Plant Physiol. 1991 Jun;96(2):363-7
PMID: 16668194
-
Interest in and limits to the utilization of reporter genes for the analysis of transcriptional regulation of nitrate reductase.
Mol Gen Genet. 1992 Nov;235(2-3):259-68
PMID: 1334527
-
Molecular cloning and characterisation of the two homologous genes coding for nitrate reductase in tobacco.
Mol Gen Genet. 1989 Mar;216(1):10-5
PMID: 2733690
-
Nitrate Reductase mRNA Regulation in Nicotiana plumbaginifolia Nitrate Reductase-Deficient Mutants.
Plant Cell. 1989 Nov;1(11):1111-1120
PMID: 12359888
-
Immunological comparisons of nitrate reductase of different plant species using monoclonal antibodies.
Plant Physiol. 1986 Jun;81(2):376-8
PMID: 16664824
-
Binary Agrobacterium vectors for plant transformation.
Nucleic Acids Res. 1984 Nov 26;12(22):8711-21
PMID: 6095209
-
Reversible light/dark modulation of spinach leaf nitrate reductase activity involves protein phosphorylation.
Arch Biochem Biophys. 1992 Jul;296(1):58-65
PMID: 1605645
-
Regulation of Corn Leaf Nitrate Reductase : II. Synthesis and Turnover of the Enzyme's Activity and Protein.
Plant Physiol. 1986 Feb;80(2):442-7
PMID: 16664640
-
Characteristics of Nicotiana tabacum nitrate reductase protein produced in Saccharomyces cerevisiae.
Biochem J. 1991 Sep 1;278 ( Pt 2):393-7
PMID: 1898332
-
Identification of the Arabidopsis CHL3 gene as the nitrate reductase structural gene NIA2.
Plant Cell. 1991 May;3(5):461-71
PMID: 1840922
-
Regulation of spinach-leaf nitrate reductase by reversible phosphorylation.
Biochim Biophys Acta. 1992 Oct 6;1137(1):121-6
PMID: 1327151
-
Rapid Modulation of Spinach Leaf Nitrate Reductase by Photosynthesis : II. In Vitro Modulation by ATP and AMP.
Plant Physiol. 1991 Jun;96(2):368-75
PMID: 16668195
-
Expression of leaf nitrate reductase genes from tomato and tobacco in relation to light-dark regimes and nitrate supply.
Plant Physiol. 1988 Oct;88(2):383-8
PMID: 16666313
-
A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia.
EMBO J. 1985 Sep;4(9):2159-65
PMID: 2866954
-
Constitutive expression of nitrate reductase allows normal growth and development of Nicotiana plumbaginifolia plants.
EMBO J. 1991 May;10(5):1027-35
PMID: 2022181
-
Purification of nitrate reductase from Nicotiana plumbaginifolia by affinity chromatography using 5'AMP-sepharose and monoclonal antibodies.
Eur J Biochem. 1989 Feb 15;179(3):617-20
PMID: 2920729