Coregulator recruitment and histone modifications in transcriptional regulation by the androgen receptor.
We have used chromatin immunoprecipitation (ChIP) assay to follow transcription factor loading and monitor changes in covalent histone modifications associated with the prostate-specific antigen and kallikrein (KLK2) genes in response to androgen and antiandrogen in LNCaP cells. The dynamics of testosterone (T)-induced loading of androgen receptor (AR) onto the proximal promoters of the genes differed significantly from that onto the distal enhancers. Significantly more holo-AR was loaded onto the enhancers than the promoters, but the receptor's residence time was more transient on the enhancers. Even though holo-AR recruited some RNA polymerase II (Pol II) onto the enhancers, the principal Pol II transcription complex was assembled on the promoters. The pure antiandrogen bicalutamide (CDX) complexed to AR elicited occupancy of the prostate-specific antigen promoter, but not that of the enhancer, whereas the partial antagonists cyproterone acetate (CPA) and mifepristone (RU486) were capable of promoting AR loading also onto the enhancer. In contrast to the CDX-occupied receptor, both CPA- and RU486-bound AR recruited Pol II and coactivators p300 and glucocorticoid receptor-interacting protein 1 (GRIP1) onto the promoter and enhancer. However, CPA and RU486 also brought about a simultaneous recruitment of the nuclear receptor corepressor (NCOR) onto the promoter as efficiently as CDX. There were dynamic changes in covalent modifications of histone H3: acetylation of lysine 9 and 14, methylation of arginine 17, phosphorylation of serine 10 as well as di- and tri-methylation at lysine 4 of the H3 N-terminal tail were enhanced in response to T, but not after CDX treatment. Collectively, these results indicate that transcriptional activation by AR is accompanied by a cascade of distinct covalent histone modifications and that the pure antiandrogen CDX and the partial antagonists CPA and RU486 exhibit clear differences in their ability to promote recruitment of histone-acetylating and histone-deacetylating complexes in human prostate cancer cells.
Biomedicum Helsinki, Institute of Biomedicine, University of Helsinki, FI-00014, Finland.
Zhigang
Z
Kang
Olli A
OA
Jänne
Jorma J
JJ
Palvimo
D000107:Acetylation; D059002:Androgen Receptor Antagonists; D000813:Anilides / Q000494:pharmacology; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D017373:Cyproterone Acetate / Q000494:pharmacology; D004257:DNA Polymerase II / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005786:Gene Expression Regulation; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008297:Male; D008745:Methylation; D015735:Mifepristone / Q000494:pharmacology; D009570:Nitriles; D009687:Nuclear Proteins / Q000378:metabolism; D051958:Nuclear Receptor Coactivator 2; D011401:Promoter Regions, Genetic / Q000235:genetics; D017430:Prostate-Specific Antigen / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism; D011944:Receptors, Androgen / Q000378:metabolism*; D020840:Tissue Kallikreins / Q000235:genetics*; D014105:Tosyl Compounds; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
D059002:Androgen Receptor Antagonists; D000813:Anilides; D006657:Histones; C498001:NCOA2 protein, human; D009570:Nitriles; D009687:Nuclear Proteins; D051958:Nuclear Receptor Coactivator 2; D011944:Receptors, Androgen; D014105:Tosyl Compounds; D015534:Trans-Activators; D014157:Transcription Factors; D015735:Mifepristone; D017373:Cyproterone Acetate; C053541:bicalutamide; D011484:Protein-Arginine N-Methyltransferases; C119966:coactivator-associated arginine methyltransferase 1; D004257:DNA Polymerase II; D020840:Tissue Kallikreins; D017430:Prostate-Specific Antigen