Seminario dal titolo “Saggi funzionali nel lievito Saccharomyces cerevisiae per lo studio di fattori di trascrizione umani e loro mutazioni associate a patologie”

Giovedì, 12 luglio 2012, ore 14:30, AULA 27, Edificio A piano terra, Area della Ricerca CNR, Pisa

05.07.2012

Giovedì, 12 luglio 2012, ore 14:30, AULA 27, Edificio A piano terra, Area della Ricerca CNR, Pisa si terrà un seminario dal titolo “Saggi funzionali nel lievito Saccharomyces cerevisiae per lo studio di fattori di trascrizione umani e loro mutazioni associate a patologie”.

Il seminario sarà tenuto dal Dott. Alberto Inga, Professore Associato del Centro Interdipartimentale di Biologia Integrata (CIBIO) dell’Università di Trento.

 

Titolo e abstract

Yeast-based assays to study human transcription factors and their disease-associated mutations.

The model organism S. cerevisiae has been used in many different settings to investigate the functions of ectopically expressed human proteins, including sequence-specific transcription factors. Starting from the analysis of the p53 transcription factor, we have recently modified a yeast-based luciferase assay to provide the opportunity for efficient investigation in a miniaturized format of several factors influencing p53 functions, including expression levels, mutations, cofactor proteins and small molecules. The system integrates i) variable expression of p53 proteins under the finely tunable GAL1,10 promoter, ii) single copy, chromosomally located p53-responsive and control luminescence reporters, iii) enhanced chemical uptake using modified ABC-transporters, iv) small-volume formats for treatment and dual-luciferase assays, and v) opportunities to co-express p53 with other cofactor proteins. The same assay format and approach has been extended to the study of other sequence-specific transcription factors, including p63 isoforms and mutations associated with ectodermal dysplasias, as well as NKX2-5 or HAND1 and mutations associated with congenital heart diseases. The main features, advantages and pitfalls of the yeast-based assays as well as recent results will be discussed.