Summary
The specific activities of the branched chain amino acyl-tRNA synthetases from the cytosolic and mitochondrial fractions ofN. crassa were low in dormant conidia and increased during germination, reaching a maximum 8 h after inoculation. This stage of development is characterised by high rates of many other cellular activities.
The increases in activity of synthetases of both cytosol and mitochondria are inhibited by cycloheximide indicating that they are synthesized on cytoplasmic ribosomes. The mitochondrial synthetases show a stimulation of their specific activity when mitochondrial RNA and protein synthesis are inhibited by either ethidium bromide or chloramphenicol suggesting that a mitochondrial translation product regulates the synthesis of the mitochondrial synthetases.
The activities of amino acyl-tRNA synthetases are dependent on energy production. When respiration is uncoupled from oxidative phosphorylation, synthetase specific activities decrease although the activities of other mitochondrial enzymes like NADH-dehydrogenase increase. This phenomenon suggests that more than one mechanism regulates the synthesis of mitochondrial proteins which are formed on cytoplasmic ribosomes.
The synthesis of branched chain amino acyl-tRNA synthetases ofNeurospora is neither repressed by their cognate amino acids, nor is there inhibition by the precursors of these amino acids, as has been observed in other amino acyl-tRNA synthetases of various organism includingNeurospora.
Similar content being viewed by others
References
Atkinson DE (1968) The energy charge of the adenylate pool as a regulatory parameter, interaction with feedback modifier. Biochemistry 7:4030–4034
Barath Z, Küntzel H (1972a) Induction of mitochondrial RNA polymerase inNeurospora crassa. Nature New Biol 240:195–197
Barath Z, Küntzel H (1972b) Cooperation of mitochondrial and nuclear genes specifying the mitochondrial genetic apparatus inNeurospora crassa. Proc Natl Acad Sci USA 69:1371–1374
Barnett WE, Brown DH, Epler JL (1967) Mitochondrial-specific amino acyl-tRNA synthetases. Proc Natl Acad Sci USA 57:1775–1781
Boguslawski G, Vodkin MH, Finkelstein DB, Fink GR (1974) Histidyl-tRNA and histidyl-tRNA synthetases in wild type and cytoplasmic petite mutants ofSaccharomyces cerevisiae. Biochemistry 13:4659–4667
Bollum FJ (1968) Filter paper disk techniques for assaying radioactive macromolecules. In: Colowick SP, Kalpan NO (eds) Methods in enzymology, vol XII-B. Academic Press, New York, pp 169–173
Brenner M, DeLorenzo F, Ames BN (1970) Energy charge and protein synthesis. Control of amino acyl-transfer ribonucleic acid synthetases. J Biol Chem 245:450–452
Buck CA, Nass MMK (1969) Studies on mitochondrial tRNA from animal cells. I. A comparison of mitochondrial and cytoplasmic tRNA and amino acyl-tRNA syntheases. J Mol Biol 41:67–82
Burkard G, Guillemant P, Weil JH (1970) Comparative studies of the tRNA's and the amino acyl-tRNA synthetases from the cytoplasmic and the chloroplast ofPhaseolus vulgaris. Biochim Biophys Acta 224:184–198
Coleman WG Jr, Williams LS (1974) First enzyme of histidine biosynthesis and repression control of histidyl-transfer ribonucleic acid synthetase ofSalmonella thyphimurium J. Bacteriol 120:390–393
Cowles JR, Key JL (1973) Demonstration of two tyrosyl-tRNA synthetases of pea roots. Biochim Biophys Acta 281:33–44
Economidis IV (1975) Studies on the regulation of the branched chain amino acyl-tRNA synthetases of the fungusNeurospora crassa. Ph D Dissertation, The University of Texas at Austin
Edwards DL, Kwiencinski F (1973) Altered mitochondrial respiration in a chromosomal mutant ofNeurospora crassa. J Bacteriol 116:610–618
Edwards DL, Rosenberg E, Maroney PA (1974) Induction of Cyanide-insensitive respiration inNeurospora crassa. J Biol Chem 249:3551–3556
Ehresmann B, Karst F, Weil JH (1971) Regulation of the biosynthesis of valyl-tRNA synthetases in yeast. Biochim Biophys Acta 254:226–236
Etten JL van, Brambl RM (1968) Protein synthesis during fungal spore germination. II. Amino acyl-soluble ribonucleic acid synthetase activities during germination ofBotryodiplodia theobromae spores. J Bacteriol 96:1042–1048
Favorova O, Kisselev L (1970) Glycyl-tRNA synthetase from rat liver: The role of tRNA in formation of glycylhydroxamates. FEBS Lett. 6:65–68
Flavell RB, Woodward DO (1970) The regulation of synthesis of Krebs cycle enzymes inNeurospora by catabolite and end product repression. Eur J Biochem 13:548–553
Greenawalt JW, Beck DP, Hawley ES (1972) Chemical and biochemical change in mitochondria during morphogenetic development ofNeurospora crassa. In: Azzone GF, Carafoli E, Lehninger AL, Quagliariello E, Siliprandi N (eds) Biochemistry and biophysics of mitochondria membrane. Academic Press, New York, pp 541–558
Harding RW, Caroline DF, Wagner RP (1970) The pyruvate dehydrogenase complex from the mitochondrial fraction ofNeurospora crassa. Arch Biochem Biophys 138:653–661
Hollomon DW (1970) Ribonucleic acid synthesis during fungal spore germination. J Gen Microbiol 62:75–87
Kisseley LL, Baturina ID (1972) Two enzymatically active forms of lysyl-tRNA synthetase forE. coli B. FEBS Lett 22:231–234
Kroon AM, Agsteribbe E, Vries H de (1972) Protein synthesis in mitochondria and chloroplasts. In: Bosch L (ed) The mechanism of protein synthesis and its regulation. North-Holland, Amsterdam, pp 540–582
Kull FJ, Jacobson KB (1969) Multiple phenylalanyl-transfer ribonucleic acid synthetase activities in the cytoplasm ofNeurospora crassa. Proc Natl Acad Sci USA 62:1137–1144
Lambowitz AM, Smith EW, Slayman CW (1972a) Electron transport inNeurospora mitochondria. Studies on wild type andPoky. J Biol Chem 247:4850–4858
Lambowitz AM, Smith EW, Slayman CW (1972b) Oxidative phosphorylation inNeurospora mitochondria. Studies on wild type,Poky and chloramphenicol—induced wild type. J Biol Chem 247:4859–4865
Leenders H, Beckers PJA (1972) The effect of changes in the respiratory metabolism upon genome activity. A correlation between reduced gene activity and an increased activity of a respiratory enzyme. J Cell Biol 55:257–265
Leenders HJ, Berendes, HD, Helmsing PJ, Derksen J, Koninkx JFJG (1974) Nuclear-mitochondrial interactions in the control of mitochondrial respiratory metabolism. Sub-Cell Biochem 3:119–147
Loftfield RB (1971) The amino acylation of transfer ribonucleic acid. In: McConey EH (ed) Protein synthesis. M. Dekker, Inc, New York, pp 1–68
Lowry OH, Rosebrough NJ, Farr, AL, Randal RH (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
McGinnis E, Williams LS (1971) Regulation of synthesis of the amino acyl-transfer ribonucleic acid synthetases for the branched chain amino acids ofEscherichia coli. J Bacteriol 108:254–262
Mirkes PE (1974) Polysomes, ribonucleic acid, and protein synthesis during germination ofNeurospora crassa Conidia J Bacteriol 117:196–202
Nazario M (1967) The accumulation of argininosuccinate inNeurospora crassa. II. Inhibition of arginyl-tRNA synthesis by argininosuccinate. Biochim Biophys Acta 145:146–152
Obrig TG, Culp WJ, McKeehan WL, Hardesty B (1971) The mechanism by which cycloheximide and related glutarimide antibiotics inhibit peptide synthesis on reticulocyte ribosomes. J Biol Chem 246:174–181
Radhakrishnan AN (1960) Preparation of an active glucose-6-phosphate dehydrogenase fromNeurospora. Biochim Biophys Acta 40:546–547
Reger BJ, Fairfield SA, Epler JL, Barnett WE (1970) Identification and origin of some chloroplast amino acyl-tRNA synthetases and tRNAs. Proc Natl Acad Sci USA 67:1207–1213
Rouget P, Chapeville F (1971) Leucyl-tRNA synthetase. Two forms of the enzyme: Relation between structural and catalytic properties. Eur J Biochem 23:459–467
Schatz G, Mason TL (1974) The biosynthesis of mitochondrial protein. Annu Rev Biochem 44:51–87
Siegel MR, Sisler HD (1964) Site of action of cycloheximide in cells ofSaccharomyces pastorianus. II. The nature of inhibition of protein synthesis in a cell-free system. Biochim Biophys Acta 87:83–89
Slayman C (1973) Adenine nucleotide levels inNeurospora, as influenced by conditions of growth and by metabolic inhibitors. J Bacteriol 114:752–766
Stuchell RN, Weinstein BI, Beattie DS (1975) Effects of ethidium bromide on the respiratory chain and oligomycin-sensitive adenosine triphosphatase in purified mitochondria from the cellular slime moldDictyostelium discoideum. J Biol Chem 250:570–576
Surdin-Kerjan Y, Cherest H, Robichon-Szulmajster H De (1973) Relationship between methionyl transfer ribonucleic acid cellular content and synthesis of methionine enzymes inSaccharomyces cerevisiae. J Bacteriol 113:1156–1160 (1973)
Vogel HJ (1956) A convenient growth medium forNeurospora (Medium N). Microb Genet Bull 13:42–43
Weeks CO, Gross SR (1971) Mutation and reversion at the Leu-5 locus ofNeurospora and its effect on the cytoplasmic and mitochondrial leucyl-tRNA synthetases. Biochem Genet 5:505–516
Westergaard O, Lindberg B (1972) An induced mitochondrial DNA polymerase fromTetrahymena. Eur J Biochem 28:422–431
Williams AL, Yem DW, McGinnis E, Williams LS (1973) Control of arginine biosynthesis inEscherichia coli: Inhibition of arginyl-transfer ribonucleic acid synthetase activity. J Bacteriol 115:228–234
Williams LS, Neidhardt FC (1969) Synthesis and inactivation of amino acyl-transfer RNA synthetase during growth ofEscherichia coli. J Mol Biol 43:529–550
Yem DW, Williams LS (1971) Inhibition of arginyl-transfer ribonucleic acid synthetase activity ofEscherichia coli by arginine biosynthetic precursors. J Bacteriol 107:589–591
Yem DW, Williams LS (1973) Evidence for the existence of two arginyl-transfer ribonucleic acid synthetase activities inEscherichia coli. J Bacteriol 113:891–894
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Economidis, I.V., Wagner, R.P. Studies on the regulation of the branched chain amino acyl-tRNA synthetases of the fungusNeurospora crassa . Wilhelm Roux' Archiv 189, 171–180 (1980). https://doi.org/10.1007/BF00868675
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00868675