Genetics, Vol 137, 659-670, Copyright © 1994

INVESTIGATIONS

The Dominant PNM2(-) Mutation Which Eliminates the {psi} Factor of Saccharomyces cerevisiae Is the Result of a Missense Mutation in the SUP35 Gene

S. M. Doel, S. J. McCready, C. R. Nierras and B. S. Cox
Present address: British Biotechnology plc, Watlington Road, Oxford, England.

The PNM2(-) mutation of Saccharomyces cerevisiae eliminates the extrachromosomal element {psi}. PNM2 is closely linked to the omnipotent suppressor gene SUP35 (also previously identified as SUP2, SUF12, SAL3 and GST1). We cloned PNM2(-) and showed that PNM2 and SUP35 are the same gene. We sequenced the PNM2(-) mutant allele and found a single G -> A transition within the N-terminal domain of the protein. We tested the effects of various constructs of SUP35 and PNM2(-) on {psi} inheritance and on allosuppressor and antisuppressor functions of the gene. We found that the C-terminal domain of SUP35 protein (SUP35p) could be independently expressed; expression produced dominant antisuppression. Disruption of the N-terminal domain of PNM2(-) destroyed the ability to eliminate {psi}. These results imply that the domains of SUP35p act in an antagonistic manner: the N-terminal domain decreases chain-termination fidelity, while the C-terminal domain imposes fidelity. Two transcripts were observed for SUP35, a major band at 2.4 kb and a minor band at 1.3 kb; the minor band corresponds to 3' sequences only. We propose a model for the function of SUP35, in which comparative levels of N- and C-terminal domains of SUP35p at the ribosome modulate translation fidelity.

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