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Effect of phosphate levels on the synthesis of acid phosphatases (EC 3.1.3.2) in Neurospora crassa

Published online by Cambridge University Press:  14 April 2009

Sergio A. Rodrigues  and
Antonio Rossi
Sergio A. Rodrigues
Affiliation:
Departamento de Química, FFCLRP-USP 14.100 Ribeirão Preto, SP, Brazil
Antonio Rossi
Affiliation:
Departamento de Química, FFCLRP-USP 14.100 Ribeirão Preto, SP, Brazil
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When grown on high-phosphate medium, the wild-type strain 74A of N. crassa synthesized two acid phosphatases, as shown by DEAE -cellulose chromatography. These purified enzymes showed heterogeneity on PAGE, low specific activities towards PNP-P, molecular weight values of at least 300000, no deviation from Michaelian behaviour, and great stability in 50 mM sodium acetate buffer, at pH 5·4, when kept at 54 °C. These acid phosphatases were synthesized in reduced amounts or not at all when the mould was grown under conditions of phosphate starvation, indicating that the level of phosphate also regulates the synthesis of the high molecular weight enzyme forms. When grown on high phosphate medium, the pho-3 mutant strain also synthesized two acid phosphatases, whose purified enzymes showed no pronounced differences when compared to those synthesized by the wild-type strain in terms of electrophoretic analysis, specific activities towards PNP-P, molecular weight values, and Michaelian behaviour. However, one enzyme form had a higher Km value and a lower heat stability than the corresponding enzyme of the wild-type strain. Even though the pho-3 locus might not be responsible for an alteration in the primary structure of the repressible acid phosphatase, it seems clear that the enzymes synthesized by the mould grown on low-or high-phosphate medium must share some structural features. Thus, the drastic differences observed in the molecular properties of the enzymes synthesized by the mould grown under conditions of phosphate starvation as opposed to phosphate repression might be due to an effect exerted by the level of inorganic phosphate in regulating the translation, post-translational modifications and/or excretion, but not necessarily the gene-directed synthesis of distinct mRNAs.

Type
Research Article
Information
Genetics Research , Volume 45 , Issue 3 , June 1985 , pp. 239 - 249
Copyright
Copyright © Cambridge University Press 1985

References

REFERENCES

Andrews, P. (1964). Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochemical Journal 91, 222–233.CrossRefGoogle ScholarPubMed
Atkinson, D. E., Hathaway, J. A. & Smith, E. C. (1965). Kinetics of regulatory enzymes. The Journal of Biological Chemistry 240, 2682–2690.CrossRefGoogle ScholarPubMed
Crocken, B. & Nyc, J. F. (1963). Utilization of L-α-glycero-phosphorylcholine by a lecithin-deficient strain of Neurospora crassa. Canadian Journal of Microbiology 9, 689–696.CrossRefGoogle Scholar
Davis, B. J. (1964). Disc-electrophoresis. II. Methods and application to human serum proteins. Annals of the New York Academy of Sciences 121, 404–427.CrossRefGoogle ScholarPubMed
Dixon, M. (1953). The determination of enzyme inhibitor constants. Biochemical Journal 55, 170–171.CrossRefGoogle ScholarPubMed
Forsthoefel, A. M. & Mishra, N. C. (1983). Biochemical genetics of Neurospora nuclease I. Isolation and characterization of nuclease (nuc) mutants. Genetical Research, Cambridge 41, 271–286.CrossRefGoogle ScholarPubMed
Free, S. J. & Metzenberg, R. L. (1982). The synthesis of alkaline phosphatase in Neurospora crassa. Biochemical Genetics 20, 883–889.CrossRefGoogle ScholarPubMed
Hartree, E. F. (1972). Determination of protein: a modification of Lowry method that gives a linear photometric response. Analytical Biochemistry 48, 422–427.CrossRefGoogle Scholar
Hasunuma, K. & Ishikawa, T. (1972). Properties of two nuclease genes in Neurospora crossa. Genetics 70, 371–384.CrossRefGoogle Scholar
Jacobs, M. M., Nyc, J. F. & Brown, D. M. (1971). Isolation and chemical properties of a repressible acid phosphatase in Neurospora crossa. The Journal of Biological Chemistry 246, 1419–1425.CrossRefGoogle Scholar
Koshland, D. E. Jr, (1970). The molecular basis for enzyme regulation. In The Enzymes (ed. Boyer, Paul D.), pp. 341–396. New York: Academic Press.Google Scholar
Kuo, M.-M. & Blumenthal, H. J. (1961). An alkaline phosphomonoesterase from Neurospora crassa. Biochimica et Biophysica Acta 54, 101–109.CrossRefGoogle ScholarPubMed
Kuo, M.-M. & Blumenthal, H. J. (1961). Purification and properties of an acid phosphomonosterase from Neurospora crassa. Biochimica et Biophysica Acta 52, 13–29.CrossRefGoogle ScholarPubMed
Levitzki, A. & Koshland, D. E. Jr. (1969). Negative cooperativity in regulatory enzymes. Proceedings of the New York Academy of Sciences 62, 1121–1128.CrossRefGoogle ScholarPubMed
Lineweaver, H. & Burk, D. (1934). The determination of enzyme dissociation constants. Journal of American Chemical Society 56, 658–666.CrossRefGoogle Scholar
Littlewood, B. S., Chia, W. & Metzenberg, R. L. (1975). Genetic control of phosphate-metabolizing enzymes in Neurospora crassa: Relationships among regulatory mutations. Genetics 79, 419–434.CrossRefGoogle ScholarPubMed
Metzenberg, R. L. (1979). Implications of some genetic control mechanisms in Neurospora. Microbiological Reviews 43, 361–383.CrossRefGoogle ScholarPubMed
Metzenberg, R. L. & Chia, W. (1979). Genetic control of phosphorus assimilation in Neurospora crossa: dose-dependent dominance and recessiveness in constitutive mutants. Genetics 93, 625–643.CrossRefGoogle Scholar
Mishra, N. C. & Forsthoefel, A. M. (1983). Biochemical genetics of Neurospora nuclease II. Mutagen sensitivity and other characteristics of the nuclease mutants. Genetical Research, Cambridge 41, 287–297.CrossRefGoogle ScholarPubMed
Nahas, E. & Rossi, A. (1984). Properties of a repressible alkaline phosphatase secreted by the wild-type strain 74A of Neurospora crossa. Phytochemistry 23, 507–510.CrossRefGoogle Scholar
Nahas, E., Terenzi, H. F. & Rossi, A. (1982). Effect of carbon source and pH on the production and secretion of acid phosphatase (EC 3.1.3.2) and alkaline phosphatase (EC 3.1.3.1) in Neurospora crossa. Journal of General Microbiology 128, 2017–2021.Google Scholar
Nelson, R. E., Lehman, J. F. & Metzenberg, R. L. (1976). Regulation of phosphate metabolism in Neurospora crossa: identification of the structural gene for repressible acid phosphatase. Genetics 84, 183–192.CrossRefGoogle Scholar
Nyc, J. F. (1967). A repressible acid phosphatase in Neurospora crassa. Biochemical and Biophysical Research Communications 27, 183–188.CrossRefGoogle ScholarPubMed
Oshima, Y. (1982). Regulatory circuits for gene expression: the metabolism of galactose and phosphate. In the molecular biology of the yeast Saccharomyces cerevisae ed. Stratherm, J. N., Jones, E. W. and Broach, J. R.), pp. 159–180. New York: Cold Spring Harbor Laboratory.Google Scholar
Stellwag, E. J. & Metzenberg, R. L. (1984). Ribosomal RNA synthesis during phosphorus starvation in wild type Neurospora and in phosphorus regulatory mutants. Molecular and General Genetics 194, 105–110.CrossRefGoogle Scholar
Vogel, H. J. (1956). A convenient growth medium for Neurospora. Microbial Genetics Bulletin 13, 42–43.Google Scholar