Abstract
The N. crassa CuMT gene has been cloned and its nucleotide sequence determined., To this end an MT specific undecanucleotide was synthesized and used for cDNA synthesis with enriched MT mRNA as a template. Sequence analysis of the cDNA obtained allowed the synthesis of a unique 21mer which was used as a hybridization probe to screen a genomic DNA library of N. crassa. Several positive clones were isolated and subjected to restriction and sequence analysis. In agreement with the published amino acid sequence, the gene codes for a polypeptide of 26 amino acid residues in length. The coding region is interrupted by a small intron. Compared to the structure of mammalian MT genes the intron-exon boundaries are located in different sequence positions. The induction of MT mRNA was studied by Northern analysis. Maximum levels of MT mRNA were detected about 1 hour after addition of copper ions to mycelium of N. crassa. The half-life time of the messenger was estimated as 2.5 hours. The CuMT amounts reach a maximum level at 3 hours after induction and thereafter remain constant.
Keywords
- Sequence Position
- Sucrose Density Gradient Centrifugation
- Cysteinyl Residue
- Rabbit Reticulocyte Lysate System
- Phosphotriester Method
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References
Lerch, K. (1980) Nature 284, 368–370.
Beltramini, M. and Lerch, K. (1983) Biochemistry 22, 2043–2048.
Vogel, N.J. (1956) Microbial Genet. Bull. 13, 42–43.
Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J. and Rutter, W.J. (1979) Biochemistry 18, 5294–5299.
Aviv, J. and Leder, P. (1972) Proc. Natl. Acad. Sci. USA 69, 1408–1412.
Noll, H. (1967) Nature 215, 360–363.
Maniatis, T., Fritsch, E.F. and Sambrook, J. (1982) Molecular cloning, Cold Spring Harbour Laboratory.
Miyoshi, K., Miyaka, T., Hozumi, T. and Itakura, K. (1980) Nucleic Acids Res. 8, 5473–5490.
Miyoshi, K., Huang, T. and Itakura, K. (1980) Nucleic Acids Res. 8, 5491–5505.
Schechtman, M.G. and Yanofsky, C. (1983) J. Mol. Appl. Genet. 2, 83–99.
Münger, K., Germann, U.A. and Lerch, K. (1985) EMBO J. 4, 2665–2668.
Maxam, A.M. and Gilbert, W. (1980) Meth. Enzymol. 65, 499–560.
Sanger, F., Nicklen, S. and Coulson, A.R. (1979) Proc. Natl. Acad. Sci. USA 74, 5463–5468.
Thomas, P.S. (1980) Proc. Natl. Acad. Sci. USA 77, 5201–5205.
Durnam, D.M. and Palmiter, R.D. (1983) Anal. Biochem 131, 385–393.
Lerch, K. and Beltramini, M. (1983) Chemica Scripta 21, 109–115.
Ohi, S., Cardenosa, G., Pine, R. and Huang, P.C. (1981) J. Biol. Chem. 256, 2180–2184.
Woudt, L.P., Pastink, A., Kempers-Veenstra, A.E., Janson, A.E.M., Mager, W.H. and Planta, R.J. (1983) Nucleis Acids Res. 11, 5347–5360.
Corden, J., Wasylyk, B., Buchwalder, A., Sassone-Corsi, P., Kedinger, C. and Chambon, P. (1980) Science 209, 1406–1414.
Breathnach, R. and Chambon, P. (1981) Annu. Rev. Biochem. 50, 349–383.
Karin, M., Haslinger, A., Holgreve, H., Richards, R.I., Krauter, P., Westphal, H.M. and Beato, M. (1984) Nature 308, 513–519.
Carter, A.D., Felber, B.K., Walling, M.-J., Jubier, M.-F., Schmidt, C.J. and Hamer, D.H. (1984) Proc. Natl. Acad. Sci. USA 81, 7392–7396.
Karin, M., Najarian, R., Haslinger, A., Valenzuela, P., Welch, J. and Fogel, S. (1984) Proc. Natl. Acad. Sci. USA 81, 337–341.
Proudfoot, N.J. and Brownlee, G.G. (1976) Nature 263, 211–214.
Zaret, K.S. and Sherman, F. (1982) Cell 28, 563–573.
Breathnach, R., Benoist, C., O’Hare, K., Gannon, F. and Chambon, P. (1978) Proc. Natl. Acad. Sci. USA 75, 4853–4857.
Langford, C.J. and Gallwitz, D. (1983) Cell 33, 519–527.
Langford, C.J., Klinz, F.-J., Donath, C. and Gallwitz, D. (1984) Cell 36, 645–653.
Glanville, N., Durnam, D.N. and Palmiter, R.D. (1981) Nature 292, 267–269.
Karin, M. and Richards, R.I. (1982) Nature 299, 797–802.
Karin, M., Haslinger, A., Holtgreve, H., Cathala, G., Slater, E. and Baxter, J.D. (1984) Cell 36, 371–379.
Durnam, D.M. and Palmiter, R.D. (1981) J. Biol. Chem. 256, 5712–5716.
Hager, L.J. and Palmiter, R.D. (1981) Nature 291, 340–342.
Karin, M. and Richards, R.I. (1984) Env. Health Persp. 54, 111–115.
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Münger, K., Germann, U.A., Lerch, K. (1987). Isolation and Regulation of Expression of the Neurospora Crassa Copper Metallothionein Gene. In: Kägi, J.H.R., Kojima, Y. (eds) Metallothionein II. Experientia Supplementum, vol 52. Birkhäuser, Basel. https://doi.org/10.1007/978-3-0348-6784-9_37
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