Newly synthesized histone H4 that is incorporated into chromatin during DNA replication is acetylated on Lysines 5 and 12. Histone deacetylases HDAC1 and HDAC2 are responsible for reducing H4 acetylation as chromatin matures. Using CRISPR/Cas9-generated hdac1 or hdac2 null fibroblasts we determined that HDAC1 and HDAC2 do not fully compensate for each other in removing de novo acetyls on H4 in vivo Proteomics of nascent chromatin and proximity ligation assays with newly replicated DNA revealed binding of ATAD2, a bromodomain-containing PTM reader that recognizes acetylated H4. ATAD2 is a transcription facilitator overexpressed in several cancers and in the SV40-transformed human fibroblast model cell line used ... More
Newly synthesized histone H4 that is incorporated into chromatin during DNA replication is acetylated on Lysines 5 and 12. Histone deacetylases HDAC1 and HDAC2 are responsible for reducing H4 acetylation as chromatin matures. Using CRISPR/Cas9-generated hdac1 or hdac2 null fibroblasts we determined that HDAC1 and HDAC2 do not fully compensate for each other in removing de novo acetyls on H4 in vivo Proteomics of nascent chromatin and proximity ligation assays with newly replicated DNA revealed binding of ATAD2, a bromodomain-containing PTM reader that recognizes acetylated H4. ATAD2 is a transcription facilitator overexpressed in several cancers and in the SV40-transformed human fibroblast model cell line used in this study. Recruitment of ATAD2 to nascent chromatin was increased in hdac2 cells over wild type, and ATAD2 depletion reduced the level of nascent chromatin-associated, acetylated H4 in wild type and hdac2 cells. We propose that overexpressed ATAD2 shifts the balance of H4 acetylation by protecting this mark from removal, and that HDAC2 but not HDAC1 can effectively compete with ATAD2 for the target acetyls. ATAD2 depletion also reduced global RNA synthesis and nascent DNA-associated RNA. Moderate dependence on ATAD2 for replication fork progression was noted only for hdac2 cells overexpressing the protein.,Copyright © 2020 American Society for Microbiology.
