TY - JOUR
T1 - Direct screening for chromatin status on DNA barcodes in yeast delineates the regulome of H3K79 methylation by Dot1
AU - Vlaming, Hanneke
AU - Molenaar, Thom M.
AU - van Welsem, Tibor
AU - Poramba-Liyanage, Deepani W.
AU - Smith, Desiree E.
AU - Velds, Arno
AU - Hoekman, Liesbeth
AU - Korthout, Tessy
AU - Hendriks, Sjoerd
AU - Maarten Altelaar, A. F.
AU - van Leeuwen, Fred
PY - 2016
Y1 - 2016
N2 - Given the frequent misregulation of chromatin in cancer, it is important to understand the cellular mechanisms that regulate chromatin structure. However, systematic screening for epigenetic regulators is challenging and often relies on laborious assays or indirect reporter readouts. Here we describe a strategy, Epi-ID, to directly assess chromatin status in thousands of mutants. In Epi-ID, chromatin status on DNA barcodes is interrogated by chromatin immunoprecipitation followed by deep sequencing, allowing for quantitative comparison of many mutants in parallel. Screening of a barcoded yeast knock-out collection for regulators of histone H3K79 methylation by Dot1 identified all known regulators as well as novel players and processes. These include histone deposition, homologous recombination, and adenosine kinase, which influences the methionine cycle. Gcn5, the acetyltransferase within the SAGA complex, was found to regulate histone methylation and H2B ubiquitination. The concept of Epi-ID is widely applicable and can be readily applied to other chromatin features.
AB - Given the frequent misregulation of chromatin in cancer, it is important to understand the cellular mechanisms that regulate chromatin structure. However, systematic screening for epigenetic regulators is challenging and often relies on laborious assays or indirect reporter readouts. Here we describe a strategy, Epi-ID, to directly assess chromatin status in thousands of mutants. In Epi-ID, chromatin status on DNA barcodes is interrogated by chromatin immunoprecipitation followed by deep sequencing, allowing for quantitative comparison of many mutants in parallel. Screening of a barcoded yeast knock-out collection for regulators of histone H3K79 methylation by Dot1 identified all known regulators as well as novel players and processes. These include histone deposition, homologous recombination, and adenosine kinase, which influences the methionine cycle. Gcn5, the acetyltransferase within the SAGA complex, was found to regulate histone methylation and H2B ubiquitination. The concept of Epi-ID is widely applicable and can be readily applied to other chromatin features.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85007315482&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/27922451
U2 - https://doi.org/10.7554/eLife.18919.001
DO - https://doi.org/10.7554/eLife.18919.001
M3 - Article
C2 - 27922451
SN - 2050-084X
VL - 5
JO - eLife
JF - eLife
IS - DECEMBER2016
M1 - e18919
ER -