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

ABSTRACT: Heterochromatin protein 1? (HP1?) interacts with chromosomes by binding to lysine 9-methylated histone H3 or DNA/RNA. HP1? is involved in various biological processes. The purpose of this study is to gain an understanding of how HP1? functions in these processes by identifying HP1?-binding proteins using mass spectrometry.We performed affinity purification of HP1?-binding proteins using G1/S phase or prometaphase HEK293T cell lysates that transiently express mock or FLAG-HP1?. Coomassie staining was performed for HP1?-binding complexes, using cell lysates prepared by affinity chromatography FLAG-agarose beads, and the bands were digested and then analyzed using a mass spectrometry.We identified 99 HP1?-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.Our results suggested that HP1? is functionally active in DNA damage response via protein-protein interaction.

SUBMITTER: Kim H 

PROVIDER: S-EPMC4720079 | biostudies-other | 2016 Jan

REPOSITORIES: biostudies-other