Fig. 4: hnRNP LL RRM2–SETD2 interaction occurs in a similar fashion as hnRNP L–SETD2 binding.
a Sequence alignment of hnRNP L RRM2 and hnRNP LL RRM2 proteins. The alignment was generated by ESPript379 with CLUSTALW80. The secondary structures of hnRNP L RRM2, as determined by DSSP, are shown above the sequences. Red squares denote identical residues whereas black triangles highlight the key residues involved in the interaction with SETD2. b, e Halo purification was performed from extracts of 293 T cells co-expressing Halo-tagged SETD2C and mCherry-HA-hnRNP L/LL constructs. Input and eluted samples were resolved on gel followed by western blotting. The experiment was repeated at least 2 times all yielding similar results. Source data are provided as a Source Data file. c Halo purification was performed from extracts of 293 T cells co-expressing Halo-HA-tagged hnRNP LL and GFP-FLAG-SETD2C constructs. Input and eluted samples were resolved on gel followed by western blotting. The experiment was repeated at least two times all yielding similar results. Source data are provided as a Source Data file. d Schematic representation of hnRNP LL segments used in purification experiments. f ITC titration data of hnRNP LL with SETD22167–2192 and its fitting curve are shown. For the arithmetic mean of KD values of the three independent experiments and the thermodynamic parameters see Supplementary Table 1. All ITC binding curves are shown in Supplementary Table 2. g (Left) Structure comparison of hnRNP L_RRM2–SETD22167–2192 complex (orange) and hnRNP LL_RRM2–SETD22167–2192 complex (purple). (Right) Two complexes are shown as ribbons with selected sidechains as sticks. KD dissociation constant, DP differential power, N binding stoichiometry. RRM–RNA recognition motif, NLS nuclear localization signal.
