Project description:Our previous report revealed that protein phosphatase 2A (PP2A), complexed with the B55delta-type regulatory subunit (i.e. Cdc55p), is solely responsible for the outstanding glycolytic activity of sake yeast strains (Watanabe et al., Appl. Environ. Microbiol. 85, e02083-18 (2019). However, how PP2A mediates yeast alcoholic fermentation remains elusive. Thus, RNA-seq analysis of S. cerevisiae cdc55-delta cells at the initial fermentation stage was performed to identify the downstream effector targeting the glycolytic control.
Project description:Total RNA samples from three replicate cultures of wild type and mutant yeast strains was isolated and expression profile done using Affymetrix arrays. Comparsion between the samples indicate how mutation in a single amino acid residue in histone H4 (H4R45H) affects gene expression in yeast. Such a mutation in histone H4 is known to generate a specific class of mutants called SWI/SNF independent (SIN) mutants, and the mutants were identified by their ability to carry out transcription in the absence of yeast chromatin remodeling complex SWI/SNF. SIN mutations are known to affect higher order chromatin structure and the comparative expression profile would help identification of genes which get affected by such altered chromatin landscape. Keywords: mutant analysis
Project description:This project aims to identify novel RNA binding proteins in the baker's yeast, Saccharomyces cerevisiae. Since interactions between RNAs and proteins may be transient, yeast cells were crosslinked with UV light at 254 nm which promotes the covalent link between proteins and RNAs. After this, polyadenylated mRNAs were purified via oligo(dT) coupled to magentic beads under stringet conditions. Finally, samples were subjected to mass spectrometry analysis. To rule out the possibility of RNA-independent binding we also analysed other samples: i) samples digested with RNase one; ii) samples where we performed competition assays with polyadenylic acid.
