Summary
The nit-2 gene of Neurospora crassa encodes a trans-acting regulatory protein that activates the expression of a number of structural genes which code for nitrogen catabolic enzymes, including nitrate reductase. The NIT2 protein contains a Cys2/Cys2-type zinc-finger DNA-binding domain that recognizes promoter regions of the Neurospora nitrogen-related genes. The NIT2 zincfinger domain/β-Gal fusion protein was shown to recognize and bind in a specific manner to two upstream fragments of the nia gene of Lycopersicon esculentum (tomato) in vitro, whereas two mutant NIT2 proteins failed to bind to the same fragments. The dissociation kinetics of the complexes formed between the NIT2 protein and the Neurospora nit-3 and the tomato nia gene promoters were examined; NIT2 binds more strongly to the nit-3 promoter DNA fragment than it does to fragments derived from the plant nitrate reductase gene itself. The observed specificity of the binding suggests the existence of a NIT2-like homolog which regulates the expression of the nitrate assimilation pathway of higher plants.
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Caboche M, Rouze P (1990) Trends Genet 6: 187–192
Caddick MX, Arst HN Jr, Taylor LH, Johnson RI, Brownlee AG (1986) EMBO J 5: 1087–1090
Casadaban MJ, Martinez-Arias A, Shapira SK, Chou J (1983) Methods Enzymol 100: 293–308
Cheng C, Dewdney J, Nam H, den Boer BGW, Goodman HM (1988) EMBO J 7: 3309–3314
Crawford NM, Campbell WH (1990) Plant Cell 2: 829–835
Daniel-Vedele F, Dorbe MF, Caboche M, Rouze P (1989) Gene 85: 371–380
Davis MA, Hynes MJ (1987) Proc Natl Acad Sci USA 84: 3753–3757
Eisen A, Taylor WE, Blumberg H, Young ET (188) Mol Cell Biol 8: 4552–4456
Evans T, Felsenfeld G (1989) Cell 58: 877–885
Faure JD, Vincentz M, Kronenberger J, Caboche M (1991) Plant J (in press)
Fried M, Crothers DM (1981) Nucleic Acids Res 9: 6505–6524
Fu Y-H, Marzluf GA (1988) J Bacteriol 170: 657–661
Fu Y-H, Marzluf GA (1990a) Mol Cell Biol 10: 1056–1065
Fu Y-H, Marzluf GA (1990b) Proc Natl Acad Sci USA 87: 5331–5335
Fu Y-H, Marzluf GA (1990c) Mol Microbiol 4: 1847–1852
Galangau F, Daniel-Vedele F, Moureaux T, Dorbe MF, Leydecker MT, Caboche M (1985) Plant Physiol 88: 383–388
Hope IA, Struhl K (1985) Cell 43: 177–188
Ko LJ, Yamamoto M, Leonard MW, George KM, Ting P, Engel JD (1991) Mol Cell Biol 11: 2778–2784
Kudla B, Caddick MX, Langdon T, Martinez-Rossi NM, Bennett CF, Sibley S, Davies RW, Arst Hn Jr (1990) EMBO J 9: 1355–1364
Lam E, Chua NH (1989) Plant Cell 1: 1147–1156
Lee H, Fu Y-H, Marzluf GA (1990) Biochemistry 29: 8779–8787
Marzluf GA (1981) Microbiol Rev 45: 437–461
Okamoto P, Fu H-Y, Marzluf GA (1991) Mol Gen Genet 227: 213–223
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Schindler U, Cashmore AR (1990) EMBO J 9: 3415–3427
Solomonson LP, Barber MJ (1990) Annu Rev Plant Physiol Plant Mol Biol 41: 225–253
Tsai SF, Martin DI, Zon LI, E'Andrea AD, Wong GG, Orkin WH (1989) Nature 339: 446–451
Trainor CD, Evans T, Felsenfeld G, Boguski MS (1990) Nature 343: 92–96
Ullmann A (1984) Gene 29: 27–31
Vincentz M, Caboche M (1991) EMBO J 10: 1027–1035
Wilson DB, Dorfman DM, Orkin SH (1990) Mol Cell Biol 10: 4854–4862
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Jarai, G., Truong, HN., Daniel-Vedele, F. et al. NIT2, the nitrogen regulatory protein of Neurospora crassa, binds upstream of nia, the tomato nitrate reductase gene, in vitro. Curr Genet 21, 37–41 (1992). https://doi.org/10.1007/BF00318652
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DOI: https://doi.org/10.1007/BF00318652