title: Lysine methylation and functional modulation of androgen receptor by Set9 methyltransferase.
abstract: Lysine methyltransferases modulate activities of transcription factors and transcription coregulators by methylating specific lysine residue(s). We report that the androgen receptor (AR) is methylated at lysine-630 by Set9, which was originally identified as a histone H3K4 monomethyltransferase. Alanine substitution of lysine-630 prevented AR methylation in vitro and in vivo. Set9 methylated the nuclear and cytoplasmic AR utilizing the cofactor S-adenosyl-methionine. A pan-methyllysine antibody recognized endogenous AR, and Set9 coimmunoprecipitated with nuclear and cytoplasmic AR. Set9 overexpression potentiated AR-mediated transactivation of the probasin promoter, whereas Set9 depletion inhibited AR activity and target gene expression. Similar to AR, chromatin occupancy of Set9 at androgen response elements (AREs) was androgen dependent, and associated with methylated histone H3K4 chromatin activation marks and p300/CBP associated factor acetyltransferase recruitment. Set9 depletion increased the histone H3K9-dimethyl repressive mark at AREs and reduced histone activation marks and occupancy of p300/CBP associated factor. K630A mutation reduced amino- and carboxy-terminal (N-C) interaction in Set9-intact cells, whereas N-C interaction for wild-type AR was reduced upon Set9 depletion. The K630A mutant was resistant to loss of activity from Set9 silencing and to increase of activity from Set9 overexpression. The K630 dependence of Set9-regulated N-C interaction and AR activity suggests that Set9 directly acts on AR at the amino acid level. Chromatin recruitment of Set9 to AREs is suggestive of its additional role as a transcriptional coactivator. Because the cellular metabolic state determines the level of S-adenosylmethionine and consequently the activity of Set9, the enhanced activity of methylated AR may have special significance in certain metabolic contexts.
authors.affiliation: Department of Molecular Medicine/Institute Biotechnology, The University of Texas Health Science Center at San Antonio, 15355 Lambda Drive, San Antonio, Texas 78245, USA.
authors.forename: Soyoung
authors.initials: S
authors.lastname: Ko
authors.forename: Jungmi
authors.initials: J
authors.lastname: Ahn
authors.forename: Chung S
authors.initials: CS
authors.lastname: Song
authors.forename: Soyoung
authors.initials: S
authors.lastname: Kim
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Knapczyk-Stwora
authors.forename: Bandana
authors.initials: B
authors.lastname: Chatterjee
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D008745:Methylation; D009154:Mutation; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism*
chemical_list: D011944:Receptors, Androgen; D011495:Histone-Lysine N-Methyltransferase; C540312:SETD7 protein, human; D008239:Lysine