The dominant PNM2- mutation which eliminates the psi factor of Saccharomyces cerevisiae is the result of a missense mutation in the SUP35 gene.
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.
Doel
S M
SM
Department of Plant Sciences, Oxford University, England.
McCready
S J
SJ
Nierras
C R
CR
Cox
B S
BS
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000235:genetics; D016386:Chromosome Walking; D005111:Extrachromosomal Inheritance; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005799:Genes, Dominant; D005800:Genes, Fungal / Q000235:genetics*; D016153:Genes, Suppressor / Q000235:genetics*; D005998:Glycine / Q000235:genetics; D008957:Models, Genetic; D010445:Peptide Elongation Factors / Q000235:genetics; D010454:Peptide Termination Factors; D010957:Plasmids / Q000235:genetics*; D017354:Point Mutation; D011328:Prions; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins; D012270:Ribosomes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
D005656:Fungal Proteins; D010445:Peptide Elongation Factors; D010454:Peptide Termination Factors; D011328:Prions; D011993:Recombinant Fusion Proteins; C068068:SUP35 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D001224:Aspartic Acid; D005998:Glycine