AMINO ACID REPLACEMENTS RESULTING FROM SUPPRESSION AND MISSENSE REVERSION OF A CHAIN-TERMINATOR MUTATION IN NEUROSPORA

T. W. Seale ¹, M. Brett ¹, A. J. Baron ¹, and J. R. S. Fincham ¹

¹ Department of Genetics, University of Leeds, Leeds, England

The Neurospora crassa super-suppressor mutation, ssu-1, suppresses the auxotrophic phenotype of the mutant am(17) by inserting tyrosine at residue 313 of NADP-specific glutamate dehydrogenase, a position occupied in the wild type by glutamate. Two classes of am(17) revertants due to further mutation within the am gene have, respectively, tyrosine and leucine at residue 313. These replacements are consistent with a chain-terminating codon in am(17) of either the amber (UAG) or the ochre type (UAA), but are inconsistent with UGA. The Leu³¹³ and Tyr³¹³ variants of the enzyme have effective activity but are grossly different from the wild type in Michaelis constants (especially for ammonium) and heat stabilities at two different pH values. They show smaller but significant differences in these respects from each other.