Abstract
The correlation between membrane phospholipid composition and total cyclic AMP levels was investigated by using Neurospora lipid auxotrophs under various supplementation conditions.
The lipid composition of the supplemented cultures was determined, and the intracellular and extracellular cyclic AMP levels were measured at various stages of the culture growth. Kinetic parameters and the thermostability of adenylate cyclase and of cyclic AMP-dependent phosphodiesterase were measured under all supplementation conditions.
In inositol deficient inl cultures the levels of intracellular cyclic AMP decreased exponentially towards the end of the log phase and thereafter. In chol-l; chol-2 cultures, grown in N-monomethylethanolamine and low choline supplementation, the level of intracellular cyclic AMP decreased as function of decreasing exogenous choline supplement. Rates of cyclic AMP extrusion in all cultures were comparable on dry weight basis, and thus not affected by the mycelial lipid composition. Adenylate cyclase activity and thermostability decreased under those supplementation conditions resulting in reduction of cyclic AMP. Cyclic AMP-dependent phosphodiesterase was insensitive to phospholipid changes.
Accordingly, it is suggested that specific perturbations in cellular phospholipid composition affect the membranebound adenylate cyclase and hence the cyclic AMP synthesis in vivo.
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Abbreviations
- PI:
-
phosphatidylinositol
- PS:
-
phosphatidylserine
- PC:
-
phosphatidylcholine
- PE:
-
phosphatidylethanolamine
- PDME:
-
phosphatidyldimethylethanolamine
- PMME:
-
phosphatidylmonomethylethanolamine
- DME:
-
N,N-dimethylethanolamine
- MME:
-
N-monomethylethanolamine
- PA:
-
phosphatidic acid
- DPG:
-
diphosphatidyl glycerol (cardiolipin)
References
Bartlett GR (1959) Phosphorus assay in column chromatography. J Biol Chem 234:466–468
Flawiá MM, Torres HN (1972) Adenylate cyclase activity in Neurospora crassa. I. General properties. J Biol Chem 247:6873–6879
Flawiá MM, Torres HN (1972a) Adenylate cyclase activity in Neurospora crassa. II. Kinetics. J Biol Chem 247:6880–6883
Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 266:497–509
Gilman AG (1970) A protein binding assay for adenosine 3′,5′-cyclic monophosphate. Proc Natl Acad Sci USA 67:305–312
Hubbard SC, Brody S (1975) Glycerophospholipid variation in choline and inositol auxotrophs of Neurospora crassa. Internal compensation among zwitterionic and anionic species. J Biol Chem 250:7173–7181
Kuksis A (1966) Newer developments in determination of bile acids and steroids by gas chromatography. In: Glick D (ed) Methods of biochemical analysis, vol 14. Interscience Publishers, New York, pp 325–454
Levey GS (1971) Restoration of glucagon responsiveness of solubilized myocardial adenyl cyclase by phosphatidylserine. Biochem Biophys Res Commun 43:108–113
Levey GS (1971a) Restoration of norephinephrine responsiveness of solubilized myocardial adenylate cyclase by phosphatidylinositol. J Biol Chem 246:7405–7407
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the Folin phenol reagent. J Biol Chem 193: 267–275
Makman RS, Sutherland EW (1965) Adenosine 3′,5′-phosphate in Escherichia coli. J Biol Chem 240:1309–1314
Matile P (1966) Inositol deficiency resulting in death: An explanation of its occurrence in Neurospora crassa. Science 151:86–88
Morris DC, Safe S, Subden RE (1974) Detection of the ergosterol and episterol isomers lichesterol and fecosterol in nystatin-resistant mutants of Neurospora crassa. Biochem Genet 12:459–466
Réthy A, Tomasi V, Trevisani A (1971) The role of lipids in the activity of adenylate cyclase of rat liver plasma membranes. Arch Biochem Biophys 147:36–40
Réthy A, Tomasi V, Trevisani A, Barnabei O (1972) The role of phosphatidylserine in the hormonal control of adenylate cyclase of rat liver plasma membranes. Biochim Biophys Acta 290:58–69
Scott WA, Solomon B (1973) Cyclic 3′,5′-AMP phosphodiesterase of Neurospora crassa. Biochem Biophys Res Commun 53:1024–1030
Scott WA, Solomon B (1975) Adenosine 3′,5′-cyclic monophosphate and morphology in Neurospora crassa: Drug-induced alterations. J Bacteriol 122:454–463
Scott WA (1976) Adenosine 3′,5′-cyclic monophosphate deficiency in Neurospora crassa. Proc Natl Acad Sci USA 73:2995–2999
Scott WA (1977) Mutations resulting in an unsaturated fatty acid requirement in Neurospora. Evidence for Δ9 defects. Biochemistry 16:5274–5280
Sokoloff L, Rothblat GH (1972) Regulation of sterol synthesis in L cells. Steady-state and transitional responses. Biochim Biophys Acta 280:172–181
Vogel HJ (1964) Distribution of lysine pathways among fungi: Evolutionary implications. Am Nat 98:435–446
White AA, Zenser TV (1971) Separation of cyclic 3′,5′-nucleotide monophosphates from other nucleotides on aluminum oxide columns. Application to the assay of adenylcyclase and gluanylcyclase. Anal Biochem 41:372–396
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Gabrielides, C., Zrike, J. & Scott, W.A. Cyclic AMP levels in relation to membrane phospholipid variations in Neurospora crassa . Arch. Microbiol. 134, 108–113 (1983). https://doi.org/10.1007/BF00407941
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DOI: https://doi.org/10.1007/BF00407941