Fig. 2: PRMT1 methylates USP11 at R433. | Nature Communications

Fig. 2: PRMT1 methylates USP11 at R433.

From: Arginine methylation and ubiquitylation crosstalk controls DNA end-resection and homologous recombination repair

Fig. 2

a Higher energy Collisional Dissociation (HCD) MS/MS spectrum and the associated peptide sequence of the arginine methylated peptide EYVELCDAAGRPDQEVAQNHK, residues 423–443 derived from USP11. b R433 is a methyl-acceptor site in cells. Autoradiograph and immunoblots of 293T cells transfected with Flag-USP11, Flag-USP11-R433K, Flag-USP11-R693K and Flag-USP11-R433K/R693K, treated with CAM/CHX and labelled with [3H]-methyl-methionine. The graph represents the quantification of n = 2 independent experiments. Data are depicted as methyl incorporation adjusted for USP11 immunoprecipitation and normalised to wild-type USP11. c USP11-R433K is the major PRMT1-mediated methylation site as determined by in vitro methylation assay. Histone H4 serves as a positive control. The graph to the right represents the quantification of n = 3 independent experiments. Data are depicted as methyl incorporation adjusted for substrate levels (USP11) and normalised to wild-type USP11 (mean ± SD). NV no value. d Protein sequence alignment by MUSCLE (multiple sequence comparison by log-expectation) using USP11 as a reference, showing conservation of R433 in several USP11 orthologues and in USP11 paralogues, USP4 and USP15. Uncropped blots and processed graphical data provided as a Source Data file.

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