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BMC Mol Biol. 2003 Feb 10;4:2. Epub 2003 Feb 10.

Viable nonsense mutants for the essential gene SUP45 of Saccharomyces cerevisiae.

Author information

  • 1Université de Rennes 1, CNRS UMR 6061, IFR 97, France. smoskalenko@hotmail.com

Abstract

BACKGROUND:

Termination of protein synthesis in eukaryotes involves at least two polypeptide release factors (eRFs) - eRF1 and eRF3. The highly conserved translation termination factor eRF1 in Saccharomyces cerevisiae is encoded by the essential gene SUP45.

RESULTS:

We have isolated five sup45-n (n from nonsense) mutations that cause nonsense substitutions in the following amino acid positions of eRF1: Y53 --> UAA, E266 --> UAA, L283 --> UAA, L317 --> UGA, E385 --> UAA. We found that full-length eRF1 protein is present in all mutants, although in decreased amounts. All mutations are situated in a weak termination context. All these sup45-n mutations are viable in different genetic backgrounds, however their viability increases after growth in the absence of wild-type allele. Any of sup45-n mutations result in temperature sensitivity (37 degrees C). Most of the sup45-n mutations lead to decreased spore viability and spores bearing sup45-n mutations are characterized by limited budding after germination leading to formation of microcolonies of 4-20 cells.

CONCLUSIONS:

Nonsense mutations in the essential gene SUP45 can be isolated in the absence of tRNA nonsense suppressors.

PMID:
12589713
[PubMed - indexed for MEDLINE]
PMCID:
PMC150568
Free PMC Article

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