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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
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

Summary

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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
Copyright
Copyright © Cambridge University Press 1985

References

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