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Plant and Cell Physiology, 1985, Vol. 26, No. 2 331-339
© 1985

Article

A Folylpolyglutamate Synthetase Lesion in the met(S2706) Mutant of Neurospora crassa

Edwin A. Cossins and Patrick Y. Chan

Department of Botany, University of Alberta Edmonton, Alberta, T6G 2E9, Canada

The met(S2706) mutant (FGSC 4248) of Neurospora crassa was cultured in Vogel's minimal medium with and without L-methionine supplementation. Methionine Stimulated growth but significant mycelial production also occurred in minimal medium. The mutant had ability to generate free and protein methionine from [35S]-cysteine in vivo but the rate of this synthesis was below that shown by a wild type strain (Lindegren A, FGSC 853). Cytosolic and mitochondrial fractions of met(S2706) also generated methionine from [3-14C]-serine. Following dialysis, this reaction was stimulated by additions of tetrahydrofolate and its diglutamate derivative. Folate analyses showed that the mutant had a folylpolyglutamate concentration that was only 10% of that detected in the wild type. Despite this, mitochondria of met(S2706) contained folates that were largely polyglutamates and the total mitochondrial folate concentration (ng/mg protein) was comparable to that of the wild type.

Assays of folylpolyglutamate synthetase showed that met(S2706) had a lesion affecting a cytosolic, tetrahydrofolate diglutamate-forming activity. Cytosolic protein had ability to catalyze a diglutamate -> hexaglutamate reaction. Mitochondria of the mutant catalyzed the formation of methylenetetrahydrofolate triglutamate from the corresponding methylene monoglutamate. It is suggested that the limited folylpolyglutamate synthesis of met(S2706) may involve cytosolic and mitochondrial folylpolyglutamate synthetase activities.

(Received September 10, 1984; Accepted December 17, 1984)
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