Summary
The sequence of an mRNA encoding nitrite reductase (NiR, EC 1.7.7.1.) from the tree Betula pendula was determined. A cDNA library constructed from leaf poly(A)+ mRNA was screened with an oligonucleotide probe deduced from NiR sequences from spinach and maize. A 2.5 kb cDNA was isolated that hybridized to an mRNA, the steady-state level of which increased markedly upon induction with nitrate. The nucleotide sequence of the cDNA contains a reading frame encoding a protein of 583 amino acids that reveals 79% identity with NiR from spinach. The transit peptide of the NiR precursor from birch was determined to be 22 amino acids in size by sequence comparison with NiR from spinach and maize and is the shortest transit peptide reported so far. A graphical evaluation of identities found in the NiR sequence alignment revealed nine well conserved sections each exceeding ten amino acids in size. Sequence comparisons with related redox proteins identified essential residues involved in cofactor binding. A putative binding site for ferredoxin was found in the N-terminal half of the protein.
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References
Aliverti A, Jansen T, Zanetti G, Ronchi S, Herrmann RG, Curti B (1990) Expression in Escherichia coli of ferredoxin-NADP+ reductase from spinach. Eur J Biochem 191:551–555
Archer EK, Keegstra K (1990) Current views on chloroplast protein important and hypotheses on the origin of the transport mechanism. J Bioenerg Biomembr 22:789–810
Back E, Burkhart W, Moyer M, Privalle L, Rothstein S (1988) Isolation of cDNA clones coding for spinach nitrite reductase: complete sequence and nitrate induction. Mol Gen Genet 212:20–26
Back E, Dunne W, Schneiderbauer A, Framond A, Rastogi R, Rothstein S (1991) Isolation of the spinach nitrite reductase gene promoter which confers nitrate inducibility on GUS gene expression in transgenic tobacco. Plant Mol Biol 17:9–18
Campbell WH (1988) Higher plant nitrate reductase: arriving at a molecular view. Curr Top Plant Biochem Physiol 7:1–15
Campbell WH, Kinghorn JR (1990) Functional domains of assimilatory nitrate reductases and nitrite reductases. TIBS 15:315–319
Friemann A, Schmitz S (1992) A new approach for displaying identities and differences among aligned amino acid sequences. Comput Appl Biosci, in press
Friemann A, Brinkmann K, Hachtel W (1991) Sequence of a cDNA encoding the bi-specific NAD(P)H nitrate reductase from the tree Betula pendula and identification of conserved protein regions. Mol Gen Genet 227:97–105
Hasumi H, Nagata E, Nakamura S (1983) Molecular heterogeneity of ferredoxin-NADP+ reductase from spinach leaves. Biochem Biophys Res Commun 110:280–286
Ingestad T (1970) A definition of optimum nutrient requirements in birch seedlings I. Physiol Plant 23:1127–1138
Ingestad T (1971) A definition of optimum nutrient requirements in birch seedlings II. Physiol Plant 24:118–125
Jofuku KD, Goldberg RB (1988) Analysis of plant gene structure. In: Shaw CH (ed) Plant molecular biology. IRL Press, Oxford, pp 37–66
Keegstra K, Olsen L, Theg S (1989) Chloroplastic precursors and their transport across the envelope membranes. Annu Rev Plant Physiol 40:471–501
Lahners K, Kramer V, Back E, Privalle L, Rothstein S (1988) Molecular cloning of complementary DNA encoding maize nitrite reductase: molecular analysis and nitrate induction. Plant Physiol 88:741–746
Lancaster JR, Vega JM, Kamin H, Orme-Johnson NR, Krueger RJ, Siegel LM (1979) Identification of the iron-sulphur centre of spinach ferredoxin-nitrite reductase as a tetranuclear centre and preliminary EPR studies of mechanism. J Biol Chem 254:1268–1272
Matsui J, Takeba G, Ida S (1990) Molecular cloning and partial amino acid sequence of rice ferredoxin-nitrite reductase. Agric Biol Chem 54:3069–3071
Ninomiya Y, Sato S (1984) A ferredoxin-like electron carrier from non-green cultured tobacco cells. Plant Cell Physiol 25:453–458
Ostrowski J, Barber MJ, Rueger DC, Miller BE, Siegel LM, Kredich NM (1989) Characterization of the flavoprotein moieties of NADPH-sulfite reductase from Salmonella typhimurium and Escherichia coli. J Biol Chem 264:15796–15808
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Siegel LM, Wilkerson JO (1989) Structure and function of spinach ferredoxin-nitrite reductase. In: Wray JL, Kinghorn JR (eds) Molecular and genetic aspects of nitrate assimilation. Oxford University Press, Oxford, pp 263–283
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stephens RM (1985) A sequencers' sequence analysis package for the IBM PC. Gene Anal Techn 2:67–75
Wray JL (1989) Molecular and genetic aspects of nitrite reduction in higher plants. In: Wray JL, Kinghorn JR (eds) Molecular and genetic aspects of nitrate assimilation. Oxford University Press, Oxford, pp 244–262
Zanetti G, Morelli D, Ronchi S, Negri A, Aliverti A, Curti B (1988) Structural studies on the interaction between ferredoxin and ferredoxin-NADP+ reductase. Biochemistry 1988:3753–3759
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Communicated by R. Hagemann
These sequence data appear in the EMBL/GenBank/DDBJ nucleotide sequence data bases under the accession number X60093
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Friemann, A., Brinkmann, K. & Hachtel, W. Sequence of a cDNA encoding nitrite reductase from the tree Betula pendula and identification of conserved protein regions. Molec. Gen. Genet. 231, 411–416 (1992). https://doi.org/10.1007/BF00292710
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DOI: https://doi.org/10.1007/BF00292710