Extended Data Figure 6: PWWP domain of DNMT3A preferentially binds to H3K36me2.
a) The dCypher approach to interrogate chromatin readers (see Methods). Biotinylatednucleosomes are immobilized on Streptavidin donor beads and GST-tagged DNMT3APWWP on glutathione AlphaLISA acceptor beads. Laser excitation of the donor generates singlet oxygen that diffuses to activate emission from the acceptor: fluorescence counts are directly proportional to the amount of donor-acceptor bridged by the nucleosome-reader interaction.
b) Quantification of AlphaLISA counts for isolated GST-DNMT3APWWP interaction titratedagainst H3K36-modified nucleosomes (me0/me1/me2/me3) from mean of two replicates.
c) ITC titration and fitting curves of human DNMT3A PWWP domain with H3.3(1-42)K36-modified peptides (me0/me1/me2/me3).
d) ITC titration and fitting curves of human DNMT3A PWWP domain with H3.1(1-42)K36-modified peptides (me0/me1/me2/me3) .