posted on 2018-08-15, 13:29authored byMingxuan Wu, Dawn Hayward, Jay H. Kalin, Yun Song, John W. R. Schwabe, Philip A. Cole
The core CoREST complex (LHC) contains histone deacetylase HDAC1 and histone demethylase LSD1 held together by the scaffold protein CoREST. Here, we analyze the purified LHC with modified peptide and reconstituted semisynthetic mononucleosome substrates. LHC demethylase activity toward methyl-Lys4 in histone H3 is strongly inhibited by H3 Lys14 acetylation, and this appears to be an intrinsic property of the LSD1 subunit. Moreover, the deacetylase selectivity of LHC unexpectedly shows a marked preference for H3 acetyl-Lys9 versus acetyl-Lys14 in nucleosome substrates but this selectivity is lost with isolated acetyl-Lys H3 protein. This diminished activity of LHC to Lys-14 deacetylation in nucleosomes is not merely due to steric accessibility based on the pattern of sensitivity of the LHC enzymatic complex to hydroxamic acid-mediated inhibition. Overall, these studies have revealed how a single Lys modification can confer a composite of resistance in chromatin to a key epigenetic enzyme complex involved in gene silencing.
Funding
We would like to thank Dirk Schwarzer (Tübingen) for helpful ideas and sharing the F-40 sortase and the globular histone H3 DNA constructs, Cynthia Wolberger (Johns Hopkins) for the core histone DNA constructs, Song Tan (Penn State) for the 146 bp 601 DNA construct, Robert Levendosky and Greg Bowman (Johns Hopkins) for the 601 DNA template and assistance with preparing the 185 bp 601 DNA. We would also like to thank Yana Li and the eukaryotic tissue culture facility (Johns Hopkins) for assistance with LHC expression. JWRS is a Wellcome Trust Senior Investigator (grant WT100237) and Royal Society Wolfson Research Merit Award Holder. He is also supported by a Biotechnology and Biological Sciences Research Council Project Grant BB/J009598/1 as well as funds from 4SC. We acknowledge NIH (GM62437), the FAMRI Foundation, and the V Foundation for financial support.
History
Citation
eLife 2018;7:e37231
Author affiliation
/Organisation/COLLEGE OF LIFE SCIENCES/Biological Sciences/Molecular & Cell Biology
Additional files
Supplementary files
. Transparent reporting form
DOI: https://doi.org/10.7554/eLife.37231.023
Data availability
All data generated or analyses during this study have been deposited in Dryad.
The following dataset was generated:
Author(s) Year Dataset title Dataset URL
Database, license,
and accessibility
information
Wu M, Dawn H, Jay
H Kalin, Yun Song,
John WR Schwabe,
Philip A Cole
2018 Data from: Lysine-14 acetylation of
histone H3 in chromatin confers
resistance to the deacetylase and
demethylase activities of an
epigenetic silencing complex
http://dx.doi.org/10.5061/dryad.413tm83
Available at Dryad
Digital Repository
under a CC0 Public
Domain Dedication