TY - JOUR
T1 - A novel 6C assay uncovers Polycomb-mediated higher order chromatin conformations
AU - Tiwari, Vijay K.
AU - Cope, Leslie
AU - McGarvey, Kelly M.
AU - Ohm, Joyce E.
AU - Baylin, Stephen B.
PY - 2008/7
Y1 - 2008/7
N2 - We describe construction of a novel modification, "6C," of chromatin looping assays that allows specific proteins that may mediate long-range chromatin interactions to be defined. This approach combines the standard looping approaches previously defined with an immunoprecipitation step to investigate involvement of the specific protein. The efficacy of this approach is demonstrated by using a Polycomb group (PcG) protein, Enhancer of Zeste (EZH2), as an example of how our assay might be used. EZH2, as a protein of the PcG complex, PRC2, has an important role in the propagation of epigenetic memory through deposition of the repressive mark, histone H3, lysine 27, tri-methylation (H3K27me3). Using our new 6C assay, we show how EZH2 is a direct mediator of long-range intra- and interchromosomal interactions that can regulate transcriptional down-regulation of multiple genes by facilitating physical proximities between distant chromatin regions, thus targeting sites within to PcG machinery.
AB - We describe construction of a novel modification, "6C," of chromatin looping assays that allows specific proteins that may mediate long-range chromatin interactions to be defined. This approach combines the standard looping approaches previously defined with an immunoprecipitation step to investigate involvement of the specific protein. The efficacy of this approach is demonstrated by using a Polycomb group (PcG) protein, Enhancer of Zeste (EZH2), as an example of how our assay might be used. EZH2, as a protein of the PcG complex, PRC2, has an important role in the propagation of epigenetic memory through deposition of the repressive mark, histone H3, lysine 27, tri-methylation (H3K27me3). Using our new 6C assay, we show how EZH2 is a direct mediator of long-range intra- and interchromosomal interactions that can regulate transcriptional down-regulation of multiple genes by facilitating physical proximities between distant chromatin regions, thus targeting sites within to PcG machinery.
UR - http://www.mendeley.com/research/novel-6c-assay-uncovers-polycombmediated-higher-order-chromatin-conformations
U2 - 10.1101/gr.073452.107
DO - 10.1101/gr.073452.107
M3 - Article
SN - 1088-9051
VL - 18
SP - 1171
EP - 1179
JO - Genome Research
JF - Genome Research
IS - 7
ER -