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DNA damage-induced chromatome analysis in yeast reveals recruitment of Ubiquitin ligases for efficient DNA repairtest1

ABSTRACT: Eukaryotic cells package their genomes around histone octamers. In response to DNA damage checkpoint kinase-induced core histone degradation leads to a 20-40% reduction in nucleosome density in yeast. To gain insights into this process we report the first comprehensive proteomic analysis of yeast chromatin and the alterations that occur in response to DNA damage. We analyzed the protein content of formaldehyde cross-linked chromatin using tandem mass tag (TMT), multiplexing and high-resolution mass spectrometry (MS), after sucrose gradient enrichment of the chromatin fraction. Quantitative damage-induced changes in the chromatin-bound proteome (called chromatome), were compared among wild-type cells and those defective for the INO80 remodeler (arp8Δ), or high mobility group box proteins (Nhp6a and Nhp6b, nhp6ΔΔ). We find massive changes in the chromatome in response to Zeocin, which are strongly attenuated in cells lacking a functional INO80 remodeler.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Culture

SUBMITTER: Jan Seebacher

LAB HEAD: Susan Gasser

PROVIDER: PXD022369 | Pride | 2021-02-02

REPOSITORIES: Pride

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