Study of genes from Xenopus laevis and Caenorhabditis elegans homologues to Saccharomyces cerevisiae SUP35, controlling ambiguity of translation in yeast

Project Information

Grant agreement ID: INTAS-93-2828

Project closed

Start date 1 June 1995

End date 31 May 1996

Funded under

• International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993-

Total cost No data

EU contribution € 50 000,00

Coordinated by

Université de Rennes 1
France

Objective

In Saccharomyces cerevisiae mutation or overexpression of the SUP35 gene leads to an omnipotent suppression of non/sense codons. In this project the expression and the functions of Xenopus laevis and Caenorhabditis elegans homologues of the yeast Saccharomyces cerevisiae SUP35 gene will be studied. These genes will be cloned, sequenced, inserted into suitable yeast expression vectors and introduced into yeast cells.

Some of the pleiotropic effects to Saccharomyces cerevisiae recessive mutations in the SUP35 gene, such as: temperature sensitivity, nonsense suppression, sensitivity to aminoglycoside antibiotic paramomycin or antimicrotubule drug benomyl, will be studied. These experiments will be performed in haploid strains carrying suppressible nonsense mutations of different types: UAA and UGA as a precondition to monitoring low level of nonsense suppression.

Diploid, heterozygous for disruption, will be transformed by the plasmids of interest and meiotic segregation will be scored after tetrad analysis.

Cell terminal phenotypes of lethal (SUP35-null allele) ascospores will be characterised for original diploid and transformed ones. This will make it possible to reveal the type of cell cycle defect and detect any slight differences in null allele behaviour in the presence of heterologous homologues. As a control to these experiments all mentioned parameters in strains transformed by Saccharomyces cerevisiae translation factor genes (TEF1 and TEF2), will be studied as nonspecific control, and Saccharomyces cerevisiae SUP35 gene and SUP35 homologue from the methylotrophic yeast Pichia methanolica (SUP35P), which was shown to complement SUP35 null mutation and capable of multicopy suppression, as a positive control. Xenopus SUP35 antibodies will be prepared in order to study the expression of the protein as has been done for SUP45, and also to study the importance of SUP35 protein for translational fidelity.

Programme(s)

• IC-INTAS - International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993-

Topic(s)

• 41 - General Biology

Call for proposal

Data not available

Funding Scheme

Data not available

Coordinator

Université de Rennes 1

• EU contribution: No data
• Address: Campus de Beaulieu ; 35042 Rennes ; France
• Total cost: No data

Participants (2)

GENOME RESEARCH LTD.

United Kingdom

• EU contribution: No data
• Address: The Wellcome Trust Genome Campus Hinxton ; CB10 1SA SAFFRON WALDEN
• Total cost: No data

St. Petersburg State University

Russia

• EU contribution: No data
• Address: 199034 St. Petersburg
• Total cost: No data