Interactome Analysis Reveals that Heterochromatin Protein 1 gamma (HP1 gamma) Is Associated with the DNA Damage Response Pathway

Abstract

Purpose Heterochromatin protein 1 gamma (HP1 gamma) interacts with chromosomes by binding to lysine 9-methylated histone H3 or DNA/RNA. HP1 gamma is involved in various biological processes. The purpose of this study is to gain an understanding of how HP1 gamma functions in these processes by identifying HP1 gamma-binding proteins using mass spectrometry. Materials and Methods We performed affinity purification of HP1 gamma-binding proteins using G(1)/S phase or prometaphase HEK293T cell lysates that transiently express mock or FLAG-HP1 gamma. Coomassie staining was performed for HP1 gamma-binding complexes, using cell lysates prepared by affinity chromatography FLAG-agarose beads, and the bands were digested and then analyzed using a mass spectrometry. Results We identified 99 HP1 gamma-binding proteins with diverse cellular functions, including spliceosome, regulation of the actin cytoskeleton, tight junction, pathogenic Escherichia coli infection, mammalian target of rapamycin signaling pathway, nucleotide excision repair, DNA replication, homologous recombination, and mismatch repair. Conclusion Our results suggested that HP1 gamma is functionally active in DNA damage response via protein-protein interaction.