IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell (1535-6108) 2009 imex curation Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 9 human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu Accepted 2010-MAR-23 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 9 imex curation Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell (1535-6108) 2009 imex curation Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 9 human-cwr_r1 Homo sapiens prostate cancer cell line CWR-R1 cwr_r1 Human prostate cancer cell line CWR-R1 confocal microscopy confocal microscopy immunostaining immunostaining Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu Accepted 2010-MAR-23 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 9 imex curation Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell (1535-6108) 2009 imex curation Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 9 human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. reChIP assay. Performed using anti-AR followed by anti-RNF6. yqiu@som.umaryland.edu Accepted 2010-MAR-23 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 9 imex curation Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell (1535-6108) 2009 imex curation Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 9 human-c4_2 Homo sapiens c4-2 androgen-independent prostate cancer cells c4_2 Human androgen-independent prostate cancer cells C4-2 This cell line is also known as C4-2B. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu Accepted 2010-MAR-23 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 9 imex curation Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell (1535-6108) 2009 imex curation Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 9 human-cwr_r1 Homo sapiens prostate cancer cell line CWR-R1 cwr_r1 Human prostate cancer cell line CWR-R1 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu Accepted 2010-MAR-23 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 9 imex curation Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell (1535-6108) 2009 imex curation Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 9 human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Xu K., Shimelis H., Linn DE., Jiang R., Yang X., Sun F., Guo Z., Chen H., Li W., Chen H., Kong X., Melamed J., Fang S., Xiao Z., Veenstra TD., Qiu Y. Cancer - Interactions investigated in the context of cancer Authors contacted regarding large amounts of missing species/source info: no reply after 14 days, so I have added what I can. yqiu@som.umaryland.edu Accepted 2010-MAR-23 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 9 imex curation andr_human Androgen receptor Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 AR DHTR NR3C4 protein protein human Homo sapiens Human MEVQLGLGRVYPRPPSKTYRGAFQNLFQSVREVIQNPGPRHPEAASAAPPGASLLLLQQQQQQQQQQQQQQQQQQQQQQQETSPRQQQQQQGEDGSPQAHRRGPTGYLVLDEEQQPSQPQSALECHPERGCVPEPGAAVAASKGLPQQLPAPPDEDDSAAPSTLSLLGPTFPGLSSCSADLKDILSEASTMQLLQQQQQEAVSEGSSSGRAREASGAPTSSKDNYLGGTSTISDNAKELCKAVSVSMGLGVEALEHLSPGEQLRGDCMYAPLLGVPPAVRPTPCAPLAECKGSLLDDSAGKSTEDTAEYSPFKGGYTKGLEGESLGCSGSAAAGSSGTLELPSTLSLYKSGALDEAAAYQSRDYYNFPLALAGPPPPPPPPHPHARIKLENPLDYGSAWAAAAAQCRYGDLASLHGAGAAGPGSGSPSAAASSSWHTLFTAEEGQLYGPCGGGGGGGGGGGGGGGGGGGGGGGEAGAVAPYGYTRPPQGLAGQESDFTAPDVWYPGGMVSRVPYPSPTCVKSEMGPWMDSYSGPYGDMRLETARDHVLPIDYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGEASSTTSPTEETTQKLTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHTQ DIP protein P10275 original sequence version: 220 DIP protein P10275-1 original sequence version: 213 DIP protein P10275 original sequence version: 216 DIP protein P10275 original sequence version: 143 DIP protein P10275 original sequence version: 174 DIP protein P10275 original sequence version: 169 DIP protein P10275 original sequence version: 188 DIP protein P10275-1 original sequence version: 188 DIP protein P10275 original sequence version: 196 DIP protein P10275 original sequence version: 217 DIP protein P10275-1 original sequence version: 217 DIP protein P10275-1 original sequence version: 207 A73432C55D39AE06 tmps2_human_gene Human TMPS2/TMPRSS2 gene gene gene human Homo sapiens Human rnf14_human E3 ubiquitin-protein ligase RNF14 RNF14 ARA54 RING finger protein 14 Androgen receptor-associated protein 54 Triad2 protein HFB30 HRIHFB2038 protein protein human Homo sapiens Human MSSEDREAQEDELLALASIYDGDEFRKAESVQGGETRIYLDLPQNFKIFVSGNSNECLQNSGFEYTICFLPPLVLNFELPPDYPSSSPPSFTLSGKWLSPTQLSALCKHLDNLWEEHRGSVVLFAWMQFLKEETLAYLNIVSPFELKIGSQKKVQRRTAQASPNTELDFGGAAGSDVDQEEIVDERAVQDVESLSNLIQEILDFDQAQQIKCFNSKLFLCSICFCEKLGSECMYFLECRHVYCKACLKDYFEIQIRDGQVQCLNCPEPKCPSVATPGQVKELVEAELFARYDRLLLQSSLDLMADVVYCPRPCCQLPVMQEPGCTMGICSSCNFAFCTLCRLTYHGVSPCKVTAEKLMDLRNEYLQADEANKRLLDQRYGKRVIQKALEEMESKEWLEKNSKSCPCCGTPIEKLDGCNKMTCTGCMQYFCWICMGSLSRANPYKHFNDPGSPCFNRLFYAVDVDDDIWEDEVED 529E3F5AF38A5DAD klk3_human Prostate-specific antigen KLK3 APS Kallikrein-3 Semenogelase Gamma-seminoprotein P-30 antigen protein protein human Homo sapiens Human MWVPVVFLTLSVTWIGAAPLILSRIVGGWECEKHSQPWQVLVASRGRAVCGGVLVHPQWVLTAAHCIRNKSVILLGRHSLFHPEDTGQVFQVSHSFPHPLYDMSLLKNRFLRPGDDSSHDLMLLRLSEPAELTDAVKVMDLPTQEPALGTTCYASGWGSIEPEEFLTPKKLQCVDLHVISNDVCAQVHPQKVTKFMLCAGRWTGGKSTCSGDSGGPLVCNGVLQGITSWGSEPCALPERPSLYTKVVHYRKWIKDTIVANP AE9E732AF872141A rnf6_human E3 ubiquitin-protein ligase RNF6 RNF6 SPG2 RING-type E3 ubiquitin transferase RNF6 protein protein human Homo sapiens Human MNQSRSRSDGGSEETLPQDHNHHENERRWQQERLHREEAYYQFINELNDEDYRLMRDHNLLGTPGEITSEELQQRLDGVKEQLASQPDLRDGTNYRDSEVPRESSHEDSLLEWLNTFRRTGNATRSGQNGNQTWRAVSRTNPNNGEFRFSLEIHVNHENRGFEIHGEDYTDIPLSDSNRDHTANRQQRSTSPVARRTRSQTSVNFNGSSSNIPRTRLASRGQNPAEGSFSTLGRLRNGIGGAAGIPRANASRTNFSSHTNQSGGSELRQREGQRFGAAHVWENGARSNVTVRNTNQRLEPIRLRSTSNSRSRSPIQRQSGTVYHNSQRESRPVQQTTRRSVRRRGRTRVFLEQDRERERRGTAYTPFSNSRLVSRITVEEGEESSRSSTAVRRHPTITLDLQVRRIRPGENRDRDSIANRTRSRVGLAENTVTIESNSGGFRRTISRLERSGIRTYVSTITVPLRRISENELVEPSSVALRSILRQIMTGFGELSSLMEADSESELQRNGQHLPDMHSELSNLGTDNNRSQHREGSSQDRQAQGDSTEMHGENETTQPHTRNSDSRGGRQLRNPNNLVETGTLPILRLAHFFLLNESDDDDRIRGLTKEQIDNLSTRHYEHNSIDSELGKICSVCISDYVTGNKLRQLPCMHEFHIHCIDRWLSENCTCPICRQPVLGSNIANNG 344584773F2E5EFC bmf_human_gene Human BMF gene gene gene human Homo sapiens Human rel1_human_gene Human REL1 gene gene gene human Homo sapiens Human klk3_human_gene Human kallikrein-related peptidase 3 (KLK3) (PSA) gene gene human Homo sapiens Human rnf6-ar 5 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 1C. The association of endogenous RNF6 with AR was enhanced by androgen treatment in the prostate cancer cell lines CWR-R1 and LNCaP. R1881, a synthetic androgen. The association of endogenous RNF6 with AR was enhanced by androgen treatment in the prostate cancer cell lines CWR-R1 and LNCaP. rnf6-ar-1 6 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Fig 1C. R1881, a synthetic androgen. In addition, the association of endogenous RNF6 with AR was enhanced by androgen treatment in the prostate cancer cell lines CWR-R1 and LNCaP. rnf6-ar-2 2 7 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig 1D. Immunofluorescence staining revealed that endogenous AR and RNF6 were colocalized in punctate structures spreading throughout in the nucleus rnf6-ar-3 5 7 unspecified role unspecified role bait bait region (possible) xlnk-n6lys-1gly N6-glycyl-L-lysine (2S)-2-amino-6-[(aminoacetyl)amino]hexanoic acid N6-(glycyl)-L-lysine N6-glycyllysine XLNK-N6Lys-1Gly CROSSLNK Glycyl lysine isopeptide (Gly-Lys) (interchain with K-...) CROSSLNK Glycyl lysine isopeptide (Lys-Gly) (interchain with G-...) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey physical association physical association Fig 3A, 3C. Consistent with our earlier observation that RNF6 induces AR ubiquitination, knockdown of endogenous RNF6 in these cells significantly diminished both basal and androgen-induced ubiquitination of AR, implying that RNF6 may modulate AR transcriptional activity via ubiquitination under both androgen-responsive and androgen-depleted conditions. AR is ubiquitinated. Knockdown of endogenous RNF6 significantly diminished both basal and androgen-induced ubiquitination of AR, implying that RNF6 may modulate AR transcriptional activity via ubiquitination under both androgen-responsive and androgen-depleted conditions. rnf6-ar-4 6 11 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region (possible) xlnk-n6lys-1gly N6-glycyl-L-lysine (2S)-2-amino-6-[(aminoacetyl)amino]hexanoic acid N6-(glycyl)-L-lysine N6-glycyllysine XLNK-N6Lys-1Gly CROSSLNK Glycyl lysine isopeptide (Gly-Lys) (interchain with K-...) CROSSLNK Glycyl lysine isopeptide (Lys-Gly) (interchain with G-...) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig 3A, S3. Consistent with our earlier observation that RNF6 induces AR ubiquitination, knockdown of endogenous RNF6 in these cells significantly diminished both basal and androgen-induced ubiquitination of AR, implying that RNF6 may modulate AR transcriptional activity via ubiquitination under both androgen-responsive and androgen-depleted conditions. Knockdown of endogenous RNF6 significantly diminished both basal and androgen-induced ubiquitination of AR. Interestingly, a dramatic increase in ubiquitination of AR was also observed in CWR-R1 and LNCaP cells treated with the synthetic androgen R1881, suggesting that ubiquitination of AR is accompanied by its activation. ar-klk3-1 5 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region (possible) xlnk-n6lys-1gly N6-glycyl-L-lysine (2S)-2-amino-6-[(aminoacetyl)amino]hexanoic acid N6-(glycyl)-L-lysine N6-glycyllysine XLNK-N6Lys-1Gly CROSSLNK Glycyl lysine isopeptide (Gly-Lys) (interchain with K-...) CROSSLNK Glycyl lysine isopeptide (Lys-Gly) (interchain with G-...) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig 3C. Knocking down endogenous RNF6 in CWR-R1 and C4-2B cells under serum-free conditions diminished the expression level of a well-established AR target gene product, prostate-specific antigen (PSA), concurrent with the reduction of AR ubiquitination. Knocking down endogenous RNF6 in CWR-R1 and C4-2B cells under serum-free conditions diminished the expression level of a well-established AR target gene product, prostate-specific antigen (PSA), concurrent with the reduction of AR ubiquitination. ar-klk3 4 11 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region (possible) xlnk-n6lys-1gly N6-glycyl-L-lysine (2S)-2-amino-6-[(aminoacetyl)amino]hexanoic acid N6-(glycyl)-L-lysine N6-glycyllysine XLNK-N6Lys-1Gly CROSSLNK Glycyl lysine isopeptide (Gly-Lys) (interchain with K-...) CROSSLNK Glycyl lysine isopeptide (Lys-Gly) (interchain with G-...) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey physical association physical association Fig 3C. Knocking down endogenous RNF6 in CWR-R1 and C4-2B cells under serum-free conditions diminished the expression level of a well-established AR target gene product, prostate-specific antigen (PSA), concurrent with the reduction of AR ubiquitination. Knocking down endogenous RNF6 diminished the expression level of a well-established AR target gene product, prostate-specific antigen (PSA), concurrent with the reduction of AR ubiquitination. ar-klk3-2 1 14 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait association association Fig 3G. the chromatin immunoprecipitation (ChIP) assays shown in Figure 3G demonstrated that knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. ChIP assays demonstrated that knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. ar-rel1 1 7 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey association association Fig 3G. Knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. Knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. ar-bmf 1 12 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait association association Fig 3G. the chromatin immunoprecipitation (ChIP) assays shown in Figure 3G demonstrated that knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. ChIP assays demonstrated that knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. ar-tmps2 1 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait association association Fig 3G. Knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. Knockdown of RNF6 in LNCaP abolished AR recruitment to a subset of AREs (including those located in the enhancer region of PSA [AREIII] and the promoter region of RLN1 and BMF) but had little effect on the other subset of AREs such as those in the promoter region of PSA (AREI/II) and TMPRSS2. rnf6-klk3 3 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey association association Fig 3H. RNF6 appeared to be detectable in the AR-containing protein complex assembled on the ARE sites in the regulatory regions of PSA (AREIII), RLN1, and BMF but not present in the complex on the ARE sites of PSA (AREI/II) and TMPRSS2. RNF6 appeared to be detectable in the AR-containing protein complex assembled on the ARE sites in the regulatory regions of PSA (AREIII), RLN1, and BMF but not present in the complex on the ARE sites of PSA (AREI/II) and TMPRSS2. rnf6-rel1 3 11 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey association association Fig 3H. RNF6 appeared to be detectable in the AR-containing protein complex assembled on the ARE sites in the regulatory regions of PSA (AREIII), RLN1, and BMF but not present in the complex on the ARE sites of PSA (AREI/II) and TMPRSS2. RNF6 appeared to be detectable in the AR-containing protein complex assembled on the ARE sites in the regulatory regions of PSA (AREIII), RLN1, and BMF but not present in the complex on the ARE sites of PSA (AREI/II) and TMPRSS2. rnf6-bmf 3 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey association association Fig 3H. RNF6 appeared to be detectable in the AR-containing protein complex assembled on the ARE sites in the regulatory regions of PSA (AREIII), RLN1, and BMF but not present in the complex on the ARE sites of PSA (AREI/II) and TMPRSS2. ar-rnf14 5 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association Fig 5A. The association between AR and endogenous ARA54 was completely abolished in CWR-R1 cells either when RNF6 was knocked down. The association between AR and endogenous ARA54 was completely abolished in CWR-R1 cells when RNF6 was knocked down. ar-rnf14-1 5 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association Fig 5A. The association between AR and endogenous ARA54 was completely abolished in CWR-R1 cells either when RNF6 was knocked down. The association between AR and endogenous ARA54 was completely abolished in CWR-R1 cells when RNF6 was knocked down.