The cycl-362 allele contains a point mutation that generates an aberrant AUG codon upstream of the normal CYC1 translation initiation codon. Mutants containing this allele express only about 2% of normal iso-1-cytochrome c, presumably due to translation initiation at the upstream AUG, termination at a UAA sequence six codons downstream, and failure to reinitiate at the normal AUG codon two nucleotides later. Both intragenic and extragenic revertants of cycl-362, expressing elevated levels of iso-1-cytochrome c, have been isolated simply by selecting for growth on lactate medium. Here we describe an improved method for isolating and readily distinguishing cis- from trans-acting suppressors of the upstream AUG. Eight different genes, designated sua1 - sua8, are represented in our current collection of extragenic suppressors; all the recessive and enhance iso-1-cytochrome c levels to 10-60% of normal. None of the sua genes is allelic to SU12 or sui3, which encode eIF-2α and eIF-2ß, respectively, or to SU11. Many of the suppressors exhibit pleiotropic phenotypes, including slow growth, cold (16°C) and heat (37°C) sensitivity. These phenotypes have been exploited to clone the SUA5, SUA7 and SUA8 genes, which are presently being characterized. The structure of cucl-362 and the number of sua genes already uncovered suggest that the SUA genes are likely to encode factors affecting several different cellular processes, including translation initiation, mRNA stability and possibly transcription start site selection.
All Science Journal Classification (ASJC) codes
- conditional mutants
- genetic suppression
- iso-1-cytochrome c
- translation initiation