Abstract
Histone acetylation is generally associated with gene activation and chromatin decondensation. Recent mass spectrometry analysis has revealed that histone H4 lysine 20, a major methylation site, can also be acetylated. To understand the function of H4 lysine 20 acetylation (H4K20ac), we have developed a specific monoclonal antibody and performed ChIP-seq analysis using HeLa-S3 cells. H4K20ac was enriched around the transcription start sites (TSSs) of minimally expressed genes and in the gene body of expressed genes, in contrast to most histone acetylation being enriched around the TSSs of expressed genes. The distribution of H4K20ac showed little correlation with known histone modifications, including histone H3 methylations. A motif search in H4K20ac-enriched sequences, together with transcription factor binding profiles based on ENCODE ChIP-seq data, revealed that most transcription activators are excluded from H4K20ac-enriched genes and a transcription repressor NRSF/REST co-localized with H4K20ac. These results suggest that H4K20ac is a unique acetylation mark associated with gene repression.
Highlights
Shown H4K20ac is present in human HeLa cells and exhibits a fast turnover[7]
RHR20KacVLR was identified in fraction (Fr) 28, separated from RHR20Kme3VLR in Fr 26 (Fig. 1b,c). This data demonstrates that H4K20ac does exist in HeLa-S3 cells, which is consistent with a recent report showing that H4K20ac is a modification generally found at low abundance, occupying about 0.3% of the total amount of acetylated lysines[7]
The degree of H4K20ac enrichment in gene bodies was much lower than that of H3K36me[3] (Supplementary Fig. S2–3b). These results suggest that the distribution of H4K20ac differs both from general histone acetylation marks, which are associated with active genes[2], and from repressive marks
Summary
Shown H4K20ac is present in human HeLa cells and exhibits a fast turnover[7]. Here, we analyzed the distribution of H4K20ac using a specific monoclonal antibody. To determine the H4K20ac localization in HeLa-S3 cells, we performed a ChIP-seq analysis using the H4K20ac-specific antibody (Fig. 2 and Supplementary Fig. S2-1a). When viewed using a genome browser together with mRNA-seq and ENCODE ChIP-seq data for H3K27me[3] and H3K27ac (typical marks for repressed and active genes, respectively), H4K20ac signals appeared to be associated with gene loci that had low or no expression of mRNA, rather than with highly transcribed genes (Fig. 2a).
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