Receptor-interacting protein 140 is a repressor of the androgen receptor activity.
The androgen receptor (AR) is a ligand-activated transcription factor that controls growth and survival of prostate cancer cells. In the present study, we investigated the regulation of AR activity by the receptor-interacting protein 140 (RIP140). We first showed that RIP140 could be coimmunoprecipitated with the receptor when coexpressed in 293T cells. This interaction appeared physiologically relevant because chromatin immunoprecipitation assays revealed that, under R1881 treatment, RIP140 could be recruited to the prostate-specific antigen encoding gene in LNCaP cells. In vitro glutathione S-transferase pull-down assays provided evidence that the carboxy-terminal domain of AR could interact with different regions of RIP140. By means of fluorescent proteins, we demonstrated that ligand-activated AR was not only able to translocate to the nucleus but also to relocate RIP140 from very structured nuclear foci to a diffuse pattern. Overexpression of RIP140 strongly repressed AR-dependent transactivation by preferentially targeting the ligand binding domain-dependent activity. Moreover, disruption of RIP140 expression induced AR overactivation, thus revealing RIP140 as a strong AR repressor. We analyzed its mechanism of transrepression and first demonstrated that different regions of RIP140 could mediate AR-dependent repression. We then showed that the carboxy-terminal end of RIP140 could reverse transcriptional intermediary factor 2-dependent overactivation of AR. The use of mutants of RIP140 allowed us to suggest that C-terminal binding protein played no role in RIP140-dependent inhibition of AR activity, whereas histone deacetylases partly regulated that transrepression. Finally, we provided evidence for a stimulation of RIP140 mRNA expression in LNCaP cells under androgen treatment, further emphasizing the role of RIP140 in androgen signaling.
Carascossa
Sophie
S
Institut National de la Santé et de la Recherche Médicale, Unité 540, 60 rue de Navacelles, F-34090 Montpellier, France.
Gobinet
Jérôme
J
Georget
Virginie
V
Lucas
Annick
A
Badia
Eric
E
Castet
Audrey
A
White
Roger
R
Nicolas
Jean-Claude
JC
Cavaillès
Vincent
V
Jalaguier
Stéphan
S
D048868:Adaptor Proteins, Signal Transducing; D000429:Alcohol Oxidoreductases; D059002:Androgen Receptor Antagonists; D000818:Animals; D019556:COS Cells; D002451:Cell Compartmentation; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D015741:Metribolone / Q000494:pharmacology; D009687:Nuclear Proteins / Q000378:metabolism*; D000078723:Nuclear Receptor Interacting Protein 1; D010750:Phosphoproteins / Q000378:metabolism; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects
D048868:Adaptor Proteins, Signal Transducing; D059002:Androgen Receptor Antagonists; D004268:DNA-Binding Proteins; C000627127:NRIP1 protein, human; D009687:Nuclear Proteins; D000078723:Nuclear Receptor Interacting Protein 1; D010750:Phosphoproteins; D012333:RNA, Messenger; D015741:Metribolone; D000429:Alcohol Oxidoreductases; C096794:C-terminal binding protein; D006655:Histone Deacetylases