Lorenzo Malquori

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Research project: "Investigating the function of introns in S.cerevisiae"

Our working hypothesis is that intronic RNA may be employed as a source of trans-acting regulatory information in eukaryotes, communicating sequence-specific signals via RNA-RNA and RNA-DNA base pairing interactions. We explored this possibility by examining the incidence and distribution of homologous sequences between yeast introns and other positions in the genome. As a complementary experimental approach we used a collection of yeast mutants deleted in different individual introns, generated in the group of Christine Guthrie (UCSF); we then analyzed the transcriptional profile of 60 mutants strains by microarray analysis in standard growth conditions and we analyzed their growth on several stress media. The combination of these two approaches led us to the identification of a subset of mutants which shows both transcriptional perturbations in standard conditions and impaired growth in stress conditions. We then constructed the transgenes for the expression of the missing introns and confirmed their expression and correct splicing upon yeast transformation. We are currently analyzing the phenotypes of the reconstituted mutant strains on stress media, by microarray analysis and by real-time PCR of selected genes.

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