Genome-wide analysis of H3K56ac across C. albicans genome in hyphae-inducing conditions
Genome-wide analysis of H3K56ac across C. albicans genome in hyphae-inducing conditions
C. albicans cultures were cross-linked with 1% formaldehyde for 15 minutes at room temperature with gentle shaking. The reaction was quenched with 125 mM glycine and cells wer havested by centrifugation. C. albicans wild-type strain SC5314 was grown over-weekend at 25 °C in YPD medium. After the weekend, the yeast was diluted, measured and grown overnight again at 0.1OD in YPD medium. Subsequently, yeasts were seeded in 90-mm diameter Petri dishes at a density of 5x10^5 cells/mL in 10 mL RPMI 1640 medium with L-Glutamine, and incubated at 37°C, 5% CO2 for 16 with (treated) or without (CTRL) 10 mM nicotinamide (NAM). ChIP was performed as previusly described (Mawer, J., Massen, J., Reichert, C., Grabenhorst, N., Mylonas, C., and Tessarz, P. (2021). Nhp2 is a reader of H2AQ105me and part of a network integrating metabolism with rRNA synthesis.EMBO reports, 22(10), e52435. doi: 10.15252/embr.202152435) except for cell lysis that was carried out by using a cryogenic freezer mill (SPEX SamplePrep 6970EFM Freezer/Mill, München, Germany). DNA from Input and ChIP samples was purified with Macherey-Nagel NucleoSpin Gel and PCR Clean-Up Kit. Libraries were generated from H3K56ac and input following a previously published protocol (Ford, E., Nikopoulou, C., Kokkalis, A., and Thanos, D. (2014). A method for generating highly multiplexed ChIP-seq libraries. BMC research notes, 7, 312. doi: 10.1186/1756-0500-7-312).