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. 2016 Apr 5;113(16):4326–4331. doi: 10.1073/pnas.1522372113

Fig. 2.

Fig. 2.

PRMT1 methylates RelA at R30 located within the DNA-binding L1 loop of the RHD. (A) Human RelA was analyzed by MS for the presence of methylarginine residues. Shown are MS/MS spectra of the nonmethylated (Upper) and dimethylated (Lower) peptide 21VEIIEQPKQRGM32 containing R30. (B) In vitro methylation assay with GST-PRMT1 and wild-type or mutant RelA (amino acids 1–107) in the presence of 3H-SAM. The RTK mutant harbors substitutions of R30, R33, and R35 to lysine. (C) TNFα stimulates RelA methylation at R30 in vivo. HEK 293T cells expressing myc-RelA or myc-RelA R30A were stimulated with TNFα for 8 h, and RelA methylation was analyzed by immunoprecipitation with anti-R30 asMe2, followed by immunoblotting using anti–c-myc (9E10) antibodies.