Gene expression analysis using RNA Sequencing of the Saccharomyces cerevisiae BY4741 H4-T30 and S47 mutant strains upon heat and osmotic stress

To study the relevance of the phosphorylation of H4 in residues T30 and S47, we assessed the genome-wide transcriptional response of the non-phosphorylatable H4-T30A and the phospho-mimetic H4-T30D mutants upon heat stress and the phospho-mimetic H4-S47D mutant upon osmotic stress . Results point out that H4-T30(A/D) and H4-S47D mutants showed an impaired stress responsive gene expression indicating that these specific residues play a relevant role in, respectively, heat and osmotic stress responsive transcriptional regulation. Wild-type and H4-T30(A/D) mutant strains were grown to log-phase on YPD medium at 30ºC (no stress) and then subjected to heat stress (10 min 39°C). Wild-type and H4-S47D mutant strains were grown to log-phase on YPD medium at 30ºC (no stress) and then subjected to osmotic stress (15 min 0,4M NaCl). Total RNA was isolated. Experiments were performed in triplicate.