Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1995 May;7(5):611–621. doi: 10.1105/tpc.7.5.611

Post-transcriptional regulation of nitrate reductase by light is abolished by an N-terminal deletion.

L Nussaume 1, M Vincentz 1, C Meyer 1, J P Boutin 1, M Caboche 1
PMCID: PMC160808  PMID: 7780309

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.

Full Text

The Full Text of this article is available as a PDF (2.1 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boutry M., Chua N. H. A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia. EMBO J. 1985 Sep;4(9):2159–2165. doi: 10.1002/j.1460-2075.1985.tb03910.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cherel I., Marion-Poll A., Meyer C., Rouze P. Immunological comparisons of nitrate reductase of different plant species using monoclonal antibodies. Plant Physiol. 1986 Jun;81(2):376–378. doi: 10.1104/pp.81.2.376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Galangau F., Daniel-Vedele F., Moureaux T., Dorbe M. F., Leydecker M. T., Caboche M. 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):383–388. doi: 10.1104/pp.88.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Huber J. L., Huber S. C., Campbell W. H., Redinbaugh M. G. Reversible light/dark modulation of spinach leaf nitrate reductase activity involves protein phosphorylation. Arch Biochem Biophys. 1992 Jul;296(1):58–65. doi: 10.1016/0003-9861(92)90544-7. [DOI] [PubMed] [Google Scholar]
  6. Kaiser W. M., Brendle-Behnisch E. Rapid Modulation of Spinach Leaf Nitrate Reductase Activity by Photosynthesis : I. Modulation in Vivo by CO(2) Availability. Plant Physiol. 1991 Jun;96(2):363–367. doi: 10.1104/pp.96.2.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kaiser W. M., Spill D. Rapid Modulation of Spinach Leaf Nitrate Reductase by Photosynthesis : II. In Vitro Modulation by ATP and AMP. Plant Physiol. 1991 Jun;96(2):368–375. doi: 10.1104/pp.96.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. MacKintosh C. Regulation of spinach-leaf nitrate reductase by reversible phosphorylation. Biochim Biophys Acta. 1992 Oct 6;1137(1):121–126. doi: 10.1016/0167-4889(92)90109-o. [DOI] [PubMed] [Google Scholar]
  9. Moureaux T., Leydecker M. T., Meyer C. 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):617–620. doi: 10.1111/j.1432-1033.1989.tb14591.x. [DOI] [PubMed] [Google Scholar]
  10. Pouteau S., Cherel I., Vaucheret H., Caboche M. Nitrate Reductase mRNA Regulation in Nicotiana plumbaginifolia Nitrate Reductase-Deficient Mutants. Plant Cell. 1989 Nov;1(11):1111–1120. doi: 10.1105/tpc.1.11.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Remmler J. L., Campbell W. H. Regulation of Corn Leaf Nitrate Reductase : II. Synthesis and Turnover of the Enzyme's Activity and Protein. Plant Physiol. 1986 Feb;80(2):442–447. doi: 10.1104/pp.80.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Truong H. N., Meyer C., Daniel-Vedele F. Characteristics of Nicotiana tabacum nitrate reductase protein produced in Saccharomyces cerevisiae. Biochem J. 1991 Sep 1;278(Pt 2):393–397. doi: 10.1042/bj2780393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Vaucheret H., Marion-Poll A., Meyer C., Faure J. D., Marin E., Caboche M. 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):259–268. doi: 10.1007/BF00279369. [DOI] [PubMed] [Google Scholar]
  14. Vaucheret H., Vincentz M., Kronenberger J., Caboche M., Rouzé P. Molecular cloning and characterisation of the two homologous genes coding for nitrate reductase in tobacco. Mol Gen Genet. 1989 Mar;216(1):10–15. doi: 10.1007/BF00332224. [DOI] [PubMed] [Google Scholar]
  15. Vincentz M., Caboche M. Constitutive expression of nitrate reductase allows normal growth and development of Nicotiana plumbaginifolia plants. EMBO J. 1991 May;10(5):1027–1035. doi: 10.1002/j.1460-2075.1991.tb08041.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wilkinson J. Q., Crawford N. M. Identification of the Arabidopsis CHL3 gene as the nitrate reductase structural gene NIA2. Plant Cell. 1991 May;3(5):461–471. doi: 10.1105/tpc.3.5.461. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES