Identification and characterization of JunD missense mutants that lack menin binding.

摘要: Menin, the product of MEN1 tumor suppressor gene, binds to AP1 transcription factor JunD and represses transcriptional activity. The effects human or mouse missense mutations upon menin interaction were studied by random alanine scanning mutagenesis binding region (amino acids 1–70). mutant proteins tested for in a reverse yeast two-hybrid assay, regulation AP1-reporter assays. Random identified two different that disrupted at amino acid 42 (G42E G42R). Mutation G42A generated did not affect binding, likely reflecting conserved nature this substitution. Furthermore, size exclusion chromatography co-migrated with wild type but (G42E). Alanine residues 30–55 revealed acids, P41 P44, critical menin. Mouse mutants P41A, G42R, G42E P44A failed bind also escaped menin's control over their At lower amounts transfected menin, effect on G42R was changed from repression activation, similar c-jun. In conclusion, small N-terminal mediates key difference between c-jun, component is dependent