Ski-interacting protein (SKIP) interacts with androgen receptor in the nucleus and modulates androgen-dependent transcription
© Abankwa et al.; licensee BioMed Central Ltd. 2013
Received: 20 November 2012
Accepted: 25 March 2013
Published: 8 April 2013
The androgen receptor (AR) is a member of the nuclear receptor (NR) superfamily of ligand-inducible DNA transcription factors, and is the major mediator of male sexual development, prostate growth and the pathogenesis of prostate cancer. Cell and gene specific regulation by the AR is determined by availability of and interaction with sets of key accessory cofactors. Ski-interacting protein (SKIP; SNW1, NCOA62) is a cofactor shown to interact with several NRs and a diverse range of other transcription factors. Interestingly, SKIP as part of the spliceosome is thought to link mRNA splicing with transcription. SKIP has not been previously shown to interact with the AR.
The aim of this study was to investigate whether SKIP interacts with the AR and modulates AR-dependent transcription. Here, we show by co-immunoprecipitation experiments that SKIP is in a complex with the AR. Moreover, SKIP increased 5α-dihydrotestosterone (DHT) induced N-terminal/C-terminal AR interaction from 12-fold to almost 300-fold in a two-hybrid assay, and enhanced AR ligand-independent AF-1 transactivation. SKIP augmented ligand- and AR-dependent transactivation in PC3 prostate cancer cells. Live-cell imaging revealed a fast (half-time=129 s) translocation of AR from the cytoplasm to the nucleus upon DHT-stimulation. Förster resonance energy transfer (FRET) experiments suggest a direct AR-SKIP interaction in the nucleus upon translocation.
Our results suggest that SKIP interacts with AR in the nucleus and enhances AR-dependent transactivation and N/C-interaction supporting a role for SKIP as an AR co-factor.
SNW/SKIP proteins, which include human Ski-interacting protein (SKIP; SNW1, NCOA62) and its yeast homologue the essential splicing factor Prp45 (Pr e-mRNA Processing 45) , are phylogenetically highly conserved , and important during early development [3, 4]. SKIP was identified by its interaction with the proto-oncogenes, v-Ski and c-Ski . In addition to pre-mRNA splicing in the spliceosome , SKIP appears to have multiple other functions in transcription . It acts as a transcriptional co-regulator of a number of key cellular signalling molecules, such as of CREB binding protein (CBP)/ p300, the nuclear co-repressor (N-CoR) and silencing mediator for retinoic acid and thyroid hormone receptors (SMRT) . It also interacts with a large range of DNA binding proteins, including Smad2 and 3 proteins of the TGF-β pathway , and proteins involved in MyoD and Notch signalling  and may be involved in Wnt signalling . A recent study suggests SKIP regulates the cell cycle arrest factor p21 (Cip1) and subsequent effects on p53-dependent DNA cell damage . This appeared to involve recruitment of SKIP to the p21 promoter where SKIP plays a critical role for splicing and p21 gene expression, suggesting a role of SKIP in cancer cell apoptosis.
The fundamental effects of SKIP in transcriptional regulation are supported by its co-regulatory effect on nuclear hormone receptors, including estrogen receptor , the Vitamin D receptor (VDR) and Retinoid X Receptor (RXR) [12–15], which it antagonistically regulates in association with SIRT1 . More recently SKIP was shown to associate with P-TEFb, c-myc and Menin to act on the HIV-1 promoter [17, 18].
The androgen receptor (AR) is a member of the nuclear hormone receptor superfamily that regulates male sexual development, and is a major player in the pathogenesis and progression of prostate cancer [19–22]. Upon binding to its natural ligands testosterone and 5α-dihydrotestosterone (DHT), the AR becomes phosphorylated and translocates into the nucleus, where it binds as a homodimer to canonical nuclear receptor inverted repeat DNA response elements to activate target gene transcription . A comprehensive ChIP-on-chip analysis of AR-responsive elements (ARE) on chromosomes 21 and 22 however suggests that the majority of the 90 identified binding sites are non-canonical, with isolated half-sites, head-to-head, head-to-tail and direct repeat AREs .
The AR contains distinct structural domains also found in other nuclear receptor superfamily members. The amino (N)-terminal transactivation domain, which is also termed ligand-independent activation-function-1 domain (AF-1), is followed by a DNA-binding domain (DBD) that is linked via a hinge region to a carboxy (C)-terminal ligand-binding domain (LBD), which contains the activation function-2 domain (AF-2) [25, 26]. Ligand binding induces interaction between the N-terminal and C-terminal domains of AR [27, 28]. This interdomain rearrangement slows ligand dissociation and AR degradation . Moreover, mutations in the AR AF-2 domain, which are associated with partial or complete androgen insensitivity syndrome, also abrogate N/C-interaction in vitro, suggesting that N/C-interaction is functionally important in vivo. The AR intrinsic N-terminal FXXLF (residues 23–27) and WXXLF (residues 433–437) motifs form amphipathic α-helices that stabilise the N/C-interaction, by binding to a hydrophobic pocket at the C-terminus . Importantly, the intrinsic (F/W)XXLF motifs compete with the similar, extrinsic LXXLL-motif from p160-family type I co-regulators for the hydrophobic pocket on the AR C-terminus . Examples for type 1 co-regulators are steroid receptor co-activators (SRC1 and SRC3), transcriptional intermediary factor 2 (TIF2) and amplified in breast cancer-1 (AIB1) [25, 32].
While AF-1 mediates hormone-independent constitutive transactivation when artificially isolated, the AF-2 is inactive in the absence of hormone, but required for strong, ligand-dependent activity in androgen-dependent prostate cancer cells [33–36]. This may be due to the fact that even in the absence of ligand the extended AF-1 fragment (including the DBD) assumes a similar reticular or speckled distribution to nuclear foci, as the full-length receptor with its co-regulators, while the isolated AF-2 is distributed homogenously in the nucleoplasm, even if ligand is added . This speckle-pattern distribution is typical for markers of various nuclear compartments, such as speckles, nucleoli and cajal bodies [38, 39]. Nuclear speckles are sites of pre-mRNA splicing and SKIP was found among other splicing proteins enriched in nuclear speckles .
In this study, we demonstrate that SKIP interacts with the AR. We show that SKIP acts in multiple ways, by augmenting AR AF-1-dependent activity as a classical type I co-activator, while it also enhanced AR N/C-interaction and AR-dependent transcription in prostate cancer cells.
Mammalian one-hybrid data show that SKIP augments ligand-independent AR transcription
SKIP facilitates interaction of AR N- and C-termini
AR transcriptional activity is modulated by ligand-mediated interdomain interaction of its N- and C-termini [27, 30]. To address, whether SKIP facilitates AR N- and C-terminal, ligand-dependent interaction, the mammalian two-hybrid system was used [43, 44]. In the mammalian two-hybrid system, potentially interacting protein fragments are fused to either the GAL4 DNA binding domain (GAL4-DBD) or the V16 transactivation domain. Interaction of the fused proteins brings the GAL4-DBD and V16 domain into a complex, which increases transcription of luciferase from a GAL4 responsive promoter. Residues 1–538 of AR were fused to the VP16 activation domain and C-terminal AR-LBD residues 642–917 to the GAL4 DNA binding domain (AR642-917) to detect N/C-terminal interaction. Modulation of this interaction by SKIP would alter reporter gene expression in HEK293.
SKIP increases AR-dependent transcription in prostate cancer cell lines
These results suggest that SKIP augments DHT-induced MMTV- and PSA-reporter activity several-fold, with a response pattern that depends on the reporter gene construct.
SKIP co-immunoprecipitates with androgen receptor
We co-expressed AR with SKIP in COS-7 cells and precipitated bound proteins from the cell lysates using anti-AR or anti-HA antibodies (Figure 4). HA tagged SKIP proteins (~62 kD) were specifically detected in AR-immunoprecipitates in similar amounts, irrespective of whether cells were DHT treated (Figure 4 lanes 1 and 2). Consistent with AR co-immunoprecipitating HA-SKIP, the inverse experiment showed that AR protein (~110 kD) was specifically detected in HA-SKIP immunoprecipitates (Figure 4 lane 6 and 7).
In conclusion, these data suggest that AR and SKIP interact in a specific complex.
FRET experiments reveal interaction of AR and SKIP in the nucleus
Additional file 2:The video shows the donor- (left, green), calculated FRET- (middle, colour coded), and the acceptor-images (right, yellow). DHT was added in frame 1 (time given in middle panel) at 10 nM. The donor images show a DHT induced increase in AR-ECFP translocation from the cytoplasm to the EYFP-SKIP positive nuclear speckles. This leads to a transient increase in FRET on the speckles, suggesting an increased number of AR/SKIP interactions. The FRET signal subsequently decreased, as the signal of EYFP-SKIP diminished. (MOV 1 MB)
In conclusion, FRET-experiments provide additional evidence that a direct and transient interaction of AR and SKIP occurs within the nucleus, which increases within minutes after DHT-stimulated nuclear translocation of AR.
In this study we provide evidence that SKIP acts as a co-regulator of AR transcription. Using mammalian one- and two-hybrid-experiments, we showed that SKIP augments AR AF-1 ligand-independent transcriptional activity and AR ligand-dependent AF-1 and AF-2 (AR N/ C-terminal) transcriptional interaction. Moreover, SKIP augmented AR-dependent transcription with two ARE-containing reporters in prostate cancer PC3 cells. Immunoprecipitation and FRET-experiments showed that AR interacts with SKIP and that the AR translocates to the nucleus within minutes after ligand stimulation to interact with SKIP.
FRET detects proximities of 3–7 nm for ECFP/EYFP-FRET pairs , which is a distance that can in most cases be regarded to report on direct interactions, considering that the size of the fluorescent proteins is already 2 nm × 4 nm . The observed high FRET efficiency between AR-ECFP/ EYFP-SKIP on the speckle patterned SKIP-positive structures is therefore in agreement with a direct interaction of these proteins.
Our FRET data are supported by our one- and two-hybrid data, which suggest that SKIP can activate both the N- and C-terminus of AR. Therefore, FRET-fluorophores on the C-terminus of AR and N-terminus of SKIP may come into very close proximity, despite the relatively large size of both proteins. The strong increase of FRET upon DHT-stimulated nuclear translocation of AR-ECFP to EYFP-SKIP positive structures that resemble nuclear speckles, can be explained by the fact that an excess of the acceptor, EYFP-SKIP, leads to high FRET-levels in a molecular complex, where the donor-acceptor-ratio is 1: >1 .
Nuclear speckles form a functional compartment within the nucleus that include splicing factors. Active gene transcription coupled with pre-mRNA splicing is thought to occur at the periphery of the splicing factor compartment [49–51]. It was therefore intriguing to see that the FRET between SKIP and AR increased from the outside to the inside of the speckle-like SKIP-positive structures. However, as fluorescently tagged proteins had to be overexpressed, care must be taken when relating the magnitude of the FRET with the affinity of the two proteins under physiological conditions. On the other hand, overexpression does not seem to induce random interaction in our experiments, as the interaction only significantly increased a) after DHT-stimulation and b) involving a translocation from the cytoplasm to the nucleus. We cannot say, whether AR is in addition targeted to other nuclear compartments.
It is of interest that the AR AF-1 domain has been shown to interact within nuclear speckles with another splicing factor ANT-1 (homologous to yeast splicing factor Prp6p) . This distribution resembles the AR-SKIP co-localization that we have observed (Figure 5). Thus AR interaction with SKIP and SKIP enhancement of AR AF-1 transcriptional activity is similar to AR/ ANT-1-dependent augmentation of transcription and splicing. Therefore, further studies will be required to determine if SKIP, like ANT-1 may recruit AR into a transcription-splicing coupled machinery.
Interestingly, the SKIP-EYFP signal disappeared completely ~16 minutes after DHT mediated translocation of AR to the nucleus, suggesting AR-mediated degradation or masking of SKIP. Loss of the SKIP-EYFP signal started soon after the maximum FRET was reached, raising the possibility that a regulated degradation is triggered at high AR concentrations. The AR has been previously linked to proteasome-mediated degradation, by its interaction with ubiquitin protease USP10 , the ubiquitin ligase ARNIP  and sensitivity of AR transactivation to proteasome inhibitors .
AR transcriptional activity is dependent on its N-terminal and C-terminal domains, which if artificially isolated can act in a ligand-independent (AF-1) and ligand-dependent (AF-2) manner, respectively [23, 34]. Immunoprecipitation and FRET-data allow us to explain the transactivation data by the direct interaction of SKIP with AR, however details of the exact mechanism are still unclear. We showed that the AR N-terminal domain was sufficient to engage SKIP to increase transactivation. It is possible that in a transcriptionally active complex, such as that induced by the N-terminal AR fragment, SKIP is engaged in the recruitment of general co-regulators such as N-CoR and p300 . In line with the latter interpretation, it was previously found that AR ligand-dependently interacts with the nuclear receptor co-repressor, N-CoR , which also interacts with SKIP .
Moreover, our two-hybrid data indicate that SKIP bi-functionally interacts with the N- and C-termini of AR, or at least facilitates this N/C- interaction. Interestingly, Saitoh et al.  observed something similar for the effect of the co-regulator CBP, which is also required for nuclear foci or speckle formation. In addition, SRC1 also modulates N/C-terminal interactions , however, this and its co-regulator activities are handled by distinct parts of the protein with an apparently higher significance for its LXXLL-motif independent interaction . Moreover, co-regulators were shown to modulate the N/C-interaction, in a cell type dependent manner. Correspondingly, their effect on the N/C-interaction did not always correlate with their effects on AR-mediated transactivation or cell growth, which may highlight the importance of specific microenvironments that influence this interaction . This may be explained by the assumption that the AF-1 may be the major transactivator under normal physiological conditions, while type I co-regulators come into play, if their concentration is high, such as observed in recurrent prostate cancer .
In conclusion, our data suggest SKIP as a novel AR co-activator that enhances its AF-1 function and interdomain-interaction, as well as AR-dependent transcription. The diverse network of interactions shown for SKIP, in particular with nuclear hormone receptors, explains its impact on several signalling pathways involved in growth and development, including possibly in AR-dependent prostate cancer cell growth.
Reporter gene assays
HEK293 cells (origin: H. sapiens: embryonal kidney cells); PC3 (origin: H. sapiens: prostate cancer cells) were maintained in medium with 10% foetal calf serum and plated 24–36 hours before transfection into 24 well plates at a density of 2×104 cells (HEK293) or 2.5×104 cells (PC3) per well so that at time of transfection cells were about 75% confluent. Cells were transfected overnight with Fugene 6 (Roche) following the manufacturer’s instructions, using 1.5 μl Fugene per 0.6 μg of total plasmid DNA per well. The ligand DHT in DMEM/F12 for HEK293 cells of RPMI for PC3 cells was added to medium supplemented with 2% charcoal stripped FCS for 24 hours. Medium was removed and cells were washed once with ice cold PBS and then lysed with 2× Promega lysis buffer. Luciferase assays were performed in triplicate with the Firefly luciferase assay kit (Promega) and measured on a luminometer (Berthold LB953 Autolumat), as described previously . The PC3 cell line expresses no endogenous AR .
Co-immunoprecipitation and western blotting
Co-immunoprecipitations were performed as follows. COS-7 cells (origin: Cercopithecus aethiops, African green monkey, derived from CV-1 cells by SV40 mediated immortalization ) cultured under standard conditions in 10 cm culture dish (NUNC) were co-transfected using Lipofectamine™ 2000 (Invitrogen) with pCMV-AR and/or SKIP-pCGN plasmid DNA (10 μg each) and cultured in the presence or absence of 1 nM DHT. Cells were harvested 48 h after transfection and were lysed in 0.5 ml of cold RIPA buffer (50 mM Tris, pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 0.5% deoxcholic acid, 1% Triton X-100) containing 1 mM PMSF and proteinase inhibitors (Roche). Lysates were sonicated for 30 seconds on ice and centrifuged to remove cell debris then pre-clarified by incubation with 40 μl of 50% protein G-Sepharose beads (Amersham) and 1 μg of rabbit IgG (Dako) at 4°C for 1 hour with slow rotation to reduce non-specific binding. After pelleting the beads, the lysates were incubated with rabbit anti-AR antibody, N20 (Santa Cruz Biotechnology), and rabbit anti-HA antibody (Santa Cruz Biotechnology), respectively, at 4°C overnight, and followed by incubating with 20 μl of 50% protein G-Sepharose beads at 4°C for 1 hour to recover AR or HA-SKIP containing protein complexes. The immunoprecipitates were washed 5 times with cold RIPA buffer and resuspended in 20 μl of sample buffer before subjecting to western blotting with antibodies to AR or HA-SKIP. Briefly, parts of the immunoprecipitated sample was resolved on an 8% SDS-PAGE, blotted and probed for HA-tagged SKIP using anti-HA antibody. The other parts were separated on gels, blotted and probed with anti-AR antibody, N20 to detect an AR N-terminal fragment. The probed blots were incubated with anti-rabbit IgG HRP (Amersham) and imaged using ECL (Amersham).
SKIP-pSG5, SKIP-pM, SKIP-pCGN were previously described [7, 8]. All mammalian one and two-hybrid plasmids (pM-GAL4DBD and pVP16AD; Clontech State, USA) and the GAL45E1bTATA-luciferase reporter  were previously described . The MMTV-luciferase reporter MMTV-luc was provided by Dr. R.M. Evans, Salk Institute, La Jolla, CA and the PSA-luciferase reporter plasmid (pGL3-PSA540-enhancer, PSA-luc) provided by Bristol-Myers Squibb (Princeton, NJ) as previously described . The vector pM-GAL4DBD-AR-AF1 expressed the activation function 1 fragment of AR (residues 1–555). The vectors pM-GAL4DBD-AR642-917 and pVP16-AR1-538 were as described . pCMV-AR WT-pCMV3.1 plasmid for co-immunoprecipitation experiment was previously described [63, 64]. All AR constructs are derived from human AR. EYFP-SKIP human Skip cDNA was amplified by PCR using primer pairs (5′-TTT GAA TTC ATG GCG CTC ACC AGC TTT TTA CCT GC-3′ and 5′-TTT GTC GAC TAT TCC TTC CTC CTC TTC TTG CC-3′), phosphorylated by T4 polynucleotide kinase, and cloned into the SmaI site of pEYFP-C1 (Clontech). The AR-pECFP-N1 plasmid was as previously described .
Donor dequenching and sensitised acceptor FRET imaging experiments
FRET experiments were performed in BHK-21 cells (origin: Mesocricetus auratus, golden hamster) that were cultured and transfected as previously described. BHK cells were chosen as they were easy to transfect and showed good expression of the constructs. Donor dequenching experiments were carried out, using a LSM 510 Meta confocal microscope (Zeiss). Fluorescent 12 bit images were recorded in the donor- (ex 405 nm, em 530–600 nm) and acceptor- (ex 514 nm, em 530–600 nm) channels before and after bleaching of EYFP (at 514 nm with 100% laser transmission at 50% output in a ROI circumscribing nuclear speckles using 200 iterations). Average bleaching of the EYFP-signal was 98.3±0.5% (sem). The average apparent donor dequenching FRET-efficiency was calculated on individual EYFP-SKIP positive speckles, as: E=1-Dbefore/Dafter, where Dbefore/after are the background corrected average donor channel signals before and after bleaching, respectively. On each recorded image 3 different SKIP-positive speckles were analysed. Sensitized acceptor emission FRET imaging was carried out essentially as described . However, due to the inherently large difference in donor and acceptor expression, only the FRET index image, FRET2 , was calculated for each time point. The colour-lookup-table was assigned to the FRET2-images using Image J. The video showing the donor, FRET and acceptor-images was generated using Image J and Quick Time Pro. The translocation index, TL, was calculated on the indicated regions using the intensity of the AR-ECFP on the speckle, S, and the AR-ECFP intensity of a reference region in the cytoplasm, C: TL=S/C. For graphical representation and curve fitting, this value was then normalized using the TL at time 0 and at the last time point 988 s: TLnorm=(TL-TL0)/(TL988-TL0). To determine the translocation half-time, τ0.5, where 50% translocation occurred, a monoexponential function was fitted: TLnorm=1-exp(−t/τ), with τ0.5= − τ ln0.5.
DA was a fellow of the Swiss National Science Foundation (PA00A-111446) and GML was a NHMRC Clinical Career Development Awardee during this study. The Adelaide Prostate Cancer Research Centre is supported by an establishment grant from the Prostate Cancer Foundation of Australia [ID 2011/0452]. WT and LB were supported by NHMRC project grant #627185.
- Figueroa JD, Hayman MJ: The human Ski-interacting protein functionally substitutes for the yeast PRP45 gene. Biochem Biophys Res Commun. 2004, 319 (4): 1105-1109. 10.1016/j.bbrc.2004.05.096.View ArticlePubMedGoogle Scholar
- Folk P, Puta F, Skruzny M: Transcriptional coregulator SNW/SKIP: the concealed tie of dissimilar pathways. Cell Mol Life Sci. 2004, 61 (6): 629-640. 10.1007/s00018-003-3215-4.View ArticlePubMedGoogle Scholar
- Kostrouchova M, Housa D, Kostrouch Z, Saudek V, Rall JE: SKIP is an indispensable factor for Caenorhabditis elegans development. Proc Natl Acad Sci USA. 2002, 99 (14): 9254-9259. 10.1073/pnas.112213799.PubMed CentralView ArticlePubMedGoogle Scholar
- Negeri D, Eggert H, Gienapp R, Saumweber H: Inducible RNA interference uncovers the Drosophila protein Bx42 as an essential nuclear cofactor involved in Notch signal transduction. Mech Dev. 2002, 117 (1–2): 151-162.View ArticlePubMedGoogle Scholar
- Dahl R, Wani B, Hayman MJ: The Ski oncoprotein interacts with Skip, the human homolog of Drosophila Bx42. Oncogene. 1998, 16 (12): 1579-1586. 10.1038/sj.onc.1201687.View ArticlePubMedGoogle Scholar
- Albers M, Diment A, Muraru M, Russell CS, Beggs JD: Identification and characterization of Prp45p and Prp46p, essential pre-mRNA splicing factors. RNA. 2003, 9 (1): 138-150. 10.1261/rna.2119903.PubMed CentralView ArticlePubMedGoogle Scholar
- Leong GM, Subramaniam N, Issa LL, Barry JB, Kino T, Driggers PH, Hayman MJ, Eisman JA, Gardiner EM: Ski-interacting protein, a bifunctional nuclear receptor coregulator that interacts with N-CoR/SMRT and p300. Biochem Biophys Res Commun. 2004, 315 (4): 1070-1076. 10.1016/j.bbrc.2004.02.004.View ArticlePubMedGoogle Scholar
- Leong GM, Subramaniam N, Figueroa J, Flanagan JL, Hayman MJ, Eisman JA, Kouzmenko AP: Ski-interacting protein interacts with Smad proteins to augment transforming growth factor-beta-dependent transcription. J Biol Chem. 2001, 276 (21): 18243-18248. 10.1074/jbc.M010815200.View ArticlePubMedGoogle Scholar
- Wang Y, Fu Y, Gao L, Zhu G, Liang J, Gao C, Huang B, Fenger U, Niehrs C, Chen YG: Xenopus skip modulates Wnt/beta-catenin signaling and functions in neural crest induction. J Biol Chem. 2010, 285 (14): 10890-10901. 10.1074/jbc.M109.058347.PubMed CentralView ArticlePubMedGoogle Scholar
- Chen Y, Zhang L, Jones KA: SKIP counteracts p53-mediated apoptosis via selective regulation of p21Cip1 mRNA splicing. Genes Dev. 2011, 25 (7): 701-716. 10.1101/gad.2002611.PubMed CentralView ArticlePubMedGoogle Scholar
- Edwards DP, Wardell SE, Boonyaratanakornkit V: Progesterone receptor interacting coregulatory proteins and cross talk with cell signaling pathways. J Steroid Biochem Mol Biol. 2002, 83 (1–5): 173-186.View ArticlePubMedGoogle Scholar
- Baudino TA, Kraichely DM, Jefcoat SC, Winchester SK, Partridge NC, MacDonald PN: Isolation and characterization of a novel coactivator protein, NCoA-62, involved in vitamin D-mediated transcription. J Biol Chem. 1998, 273 (26): 16434-16441. 10.1074/jbc.273.26.16434.View ArticlePubMedGoogle Scholar
- Zhang C, Baudino TA, Dowd DR, Tokumaru H, Wang W, MacDonald PN: Ternary complexes and cooperative interplay between NCoA-62/Ski-interacting protein and steroid receptor coactivators in vitamin D receptor-mediated transcription. J Biol Chem. 2001, 276 (44): 40614-40620. 10.1074/jbc.M106263200.View ArticlePubMedGoogle Scholar
- Zhang C, Dowd DR, Staal A, Gu C, Lian JB, van Wijnen AJ, Stein GS, MacDonald PN: Nuclear coactivator-62 kDa/Ski-interacting protein is a nuclear matrix-associated coactivator that may couple vitamin D receptor-mediated transcription and RNA splicing. J Biol Chem. 2003, 278 (37): 35325-35336. 10.1074/jbc.M305191200.View ArticlePubMedGoogle Scholar
- Barry JB, Leong GM, Church WB, Issa LL, Eisman JA, Gardiner EM: Interactions of SKIP/NCoA-62, TFIIB, and retinoid X receptor with vitamin D receptor helix H10 residues. J Biol Chem. 2003, 278 (10): 8224-8228. 10.1074/jbc.C200712200.View ArticlePubMedGoogle Scholar
- Kang MR, Lee SW, Um E, Kang HT, Hwang ES, Kim EJ, Um SJ: Reciprocal roles of SIRT1 and SKIP in the regulation of RAR activity: implication in the retinoic acid-induced neuronal differentiation of P19 cells. Nucleic Acids Res. 2010, 38 (3): 822-831. 10.1093/nar/gkp1056.PubMed CentralView ArticlePubMedGoogle Scholar
- Bres V, Gomes N, Pickle L, Jones KA: A human splicing factor, SKIP, associates with P-TEFb and enhances transcription elongation by HIV-1 Tat. Genes Dev. 2005, 19 (10): 1211-1226. 10.1101/gad.1291705.PubMed CentralView ArticlePubMedGoogle Scholar
- Bres V, Yoshida T, Pickle L, Jones KA: SKIP interacts with c-Myc and Menin to promote HIV-1 Tat transactivation. Mol Cell. 2009, 36 (1): 75-87. 10.1016/j.molcel.2009.08.015.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang Q, Li W, Zhang Y, Yuan X, Xu K, Yu J, Chen Z, Beroukhim R, Wang H, Lupien M: Androgen receptor regulates a distinct transcription program in androgen-independent prostate cancer. Cell. 2009, 138 (2): 245-256. 10.1016/j.cell.2009.04.056.PubMed CentralView ArticlePubMedGoogle Scholar
- Bennett NC, Gardiner RA, Hooper JD, Johnson DW, Gobe GC: Molecular cell biology of androgen receptor signalling. Int J Biochem Cell Biol. 2010, 42 (6): 813-827. 10.1016/j.biocel.2009.11.013.View ArticlePubMedGoogle Scholar
- Heinlein CA, Chang C: Androgen receptor in prostate cancer. Endocr Rev. 2004, 25 (2): 276-308. 10.1210/er.2002-0032.View ArticlePubMedGoogle Scholar
- Gelmann EP: Molecular biology of the androgen receptor. J Clin Oncol. 2002, 20 (13): 3001-3015. 10.1200/JCO.2002.10.018.View ArticlePubMedGoogle Scholar
- Dehm SM, Tindall DJ: Androgen receptor structural and functional elements: role and regulation in prostate cancer. Mol Endocrinol. 2007, 21 (12): 2855-2863. 10.1210/me.2007-0223.View ArticlePubMedGoogle Scholar
- Wang Q, Li W, Liu XS, Carroll JS, Janne OA, Keeton EK, Chinnaiyan AM, Pienta KJ, Brown M: A hierarchical network of transcription factors governs androgen receptor-dependent prostate cancer growth. Mol Cell. 2007, 27 (3): 380-392. 10.1016/j.molcel.2007.05.041.PubMed CentralView ArticlePubMedGoogle Scholar
- Heinlein CA, Chang C: Androgen receptor (AR) coregulators: an overview. Endocr Rev. 2002, 23 (2): 175-200. 10.1210/er.23.2.175.View ArticlePubMedGoogle Scholar
- Gao W, Bohl CE, Dalton JT: Chemistry and structural biology of androgen receptor. Chem Rev. 2005, 105 (9): 3352-3370. 10.1021/cr020456u.PubMed CentralView ArticlePubMedGoogle Scholar
- Langley E, Kemppainen JA, Wilson EM: Intermolecular NH2-/carboxyl-terminal interactions in androgen receptor dimerization revealed by mutations that cause androgen insensitivity. J Biol Chem. 1998, 273 (1): 92-101. 10.1074/jbc.273.1.92.View ArticlePubMedGoogle Scholar
- He B, Wilson EM: The NH(2)-terminal and carboxyl-terminal interaction in the human androgen receptor. Mol Genet Metab. 2002, 75 (4): 293-298. 10.1016/S1096-7192(02)00009-4.View ArticlePubMedGoogle Scholar
- Zhou ZX, Lane MV, Kemppainen JA, French FS, Wilson EM: Specificity of ligand-dependent androgen receptor stabilization: receptor domain interactions influence ligand dissociation and receptor stability. Mol Endocrinol. 1995, 9 (2): 208-218. 10.1210/me.9.2.208.PubMedGoogle Scholar
- He B, Kemppainen JA, Wilson EM: FXXLF and WXXLF sequences mediate the NH2-terminal interaction with the ligand binding domain of the androgen receptor. J Biol Chem. 2000, 275 (30): 22986-22994. 10.1074/jbc.M002807200.View ArticlePubMedGoogle Scholar
- He B, Bowen NT, Minges JT, Wilson EM: Androgen-induced NH2- and COOH-terminal Interaction Inhibits p160 coactivator recruitment by activation function 2. J Biol Chem. 2001, 276 (45): 42293-42301. 10.1074/jbc.M107492200.View ArticlePubMedGoogle Scholar
- McKenna NJ, Lanz RB, O’Malley BW: Nuclear receptor coregulators: cellular and molecular biology. Endocr Rev. 1999, 20 (3): 321-344. 10.1210/er.20.3.321.PubMedGoogle Scholar
- Tora L, White J, Brou C, Tasset D, Webster N, Scheer E, Chambon P: The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell. 1989, 59 (3): 477-487. 10.1016/0092-8674(89)90031-7.View ArticlePubMedGoogle Scholar
- Danielian PS, White R, Lees JA, Parker MG: Identification of a conserved region required for hormone dependent transcriptional activation by steroid hormone receptors. EMBO J. 1992, 11 (3): 1025-1033.PubMed CentralPubMedGoogle Scholar
- Dehm SM, Tindall DJ: Ligand-independent androgen receptor activity is activation function-2-independent and resistant to antiandrogens in androgen refractory prostate cancer cells. J Biol Chem. 2006, 281 (38): 27882-27893. 10.1074/jbc.M605002200.View ArticlePubMedGoogle Scholar
- A unified nomenclature system for the nuclear receptor superfamily. Cell. 1999, 97 (2): 161-163.
- Saitoh M, Takayanagi R, Goto K, Fukamizu A, Tomura A, Yanase T, Nawata H: The presence of both the amino- and carboxyl-terminal domains in the AR is essential for the completion of a transcriptionally active form with coactivators and intranuclear compartmentalization common to the steroid hormone receptors: a three-dimensional imaging study. Mol Endocrinol. 2002, 16 (4): 694-706. 10.1210/me.16.4.694.View ArticlePubMedGoogle Scholar
- Handwerger KE, Gall JG: Subnuclear organelles: new insights into form and function. Trends Cell Biol. 2006, 16 (1): 19-26. 10.1016/j.tcb.2005.11.005.View ArticlePubMedGoogle Scholar
- Lamond AI, Spector DL: Nuclear speckles: a model for nuclear organelles. Nat Rev Mol Cell Biol. 2003, 4 (8): 605-612. 10.1038/nrm1172.View ArticlePubMedGoogle Scholar
- Mintz PJ, Patterson SD, Neuwald AF, Spahr CS, Spector DL: Purification and biochemical characterization of interchromatin granule clusters. EMBO J. 1999, 18 (15): 4308-4320. 10.1093/emboj/18.15.4308.PubMed CentralView ArticlePubMedGoogle Scholar
- Need EF, Scher HI, Peters AA, Moore NL, Cheong A, Ryan CJ, Wittert GA, Marshall VR, Tilley WD, Buchanan G: A novel androgen receptor amino terminal region reveals two classes of amino/carboxyl interaction-deficient variants with divergent capacity to activate responsive sites in chromatin. Endocrinology. 2009, 150 (6): 2674-2682. 10.1210/en.2008-1181.PubMed CentralView ArticlePubMedGoogle Scholar
- Simental JA, Sar M, Lane MV, French FS, Wilson EM: Transcriptional activation and nuclear targeting signals of the human androgen receptor. J Biol Chem. 1991, 266 (1): 510-518.PubMedGoogle Scholar
- Fields S, Song O: A novel genetic system to detect protein-protein interactions. Nature. 1989, 340 (6230): 245-246. 10.1038/340245a0.View ArticlePubMedGoogle Scholar
- Chien CT, Bartel PL, Sternglanz R, Fields S: The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci USA. 1991, 88 (21): 9578-9582. 10.1073/pnas.88.21.9578.PubMed CentralView ArticlePubMedGoogle Scholar
- Abankwa D, Vogel H: A FRET map of membrane anchors suggests distinct microdomains of heterotrimeric G proteins. J Cell Sci. 2007, 120 (Pt 16): 2953-2962.View ArticlePubMedGoogle Scholar
- Patterson GH, Piston DW, Barisas BG: Forster distances between green fluorescent protein pairs. Anal Biochem. 2000, 284 (2): 438-440. 10.1006/abio.2000.4708.View ArticlePubMedGoogle Scholar
- Vogel SS, Thaler C, Koushik SV: Fanciful FRET. Sci STKE. 2006, 2006 (331): re2-PubMedGoogle Scholar
- Berney C, Danuser G: FRET or no FRET: a quantitative comparison. Biophys J. 2003, 84 (6): 3992-4010. 10.1016/S0006-3495(03)75126-1.PubMed CentralView ArticlePubMedGoogle Scholar
- Melcak I, Cermanova S, Jirsova K, Koberna K, Malinsky J, Raska I: Nuclear pre-mRNA compartmentalization: trafficking of released transcripts to splicing factor reservoirs. Mol Biol Cell. 2000, 11 (2): 497-510.PubMed CentralView ArticlePubMedGoogle Scholar
- Misteli T: Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function. J Cell Sci. 2000, 113 (Pt 11): 1841-1849.PubMedGoogle Scholar
- Spector DL, Lamond AI: Nuclear speckles. Cold Spring Harb Perspect Biol. 2011, 3 (2):
- Zhao Y, Goto K, Saitoh M, Yanase T, Nomura M, Okabe T, Takayanagi R, Nawata H: Activation function-1 domain of androgen receptor contributes to the interaction between subnuclear splicing factor compartment and nuclear receptor compartment. Identification of the p102 U5 small nuclear ribonucleoprotein particle-binding protein as a coactivator for the receptor. J Biol Chem. 2002, 277 (33): 30031-30039. 10.1074/jbc.M203811200.View ArticlePubMedGoogle Scholar
- Faus H, Meyer HA, Huber M, Bahr I, Haendler B: The ubiquitin-specific protease USP10 modulates androgen receptor function. Mol Cell Endocrinol. 2005, 245 (1–2): 138-146.View ArticlePubMedGoogle Scholar
- Beitel LK, Elhaji YA, Lumbroso R, Wing SS, Panet-Raymond V, Gottlieb B, Pinsky L, Trifiro MA: Cloning and characterization of an androgen receptor N-terminal-interacting protein with ubiquitin-protein ligase activity. J Mol Endocrinol. 2002, 29 (1): 41-60. 10.1677/jme.0.0290041.View ArticlePubMedGoogle Scholar
- Lin HK, Altuwaijri S, Lin WJ, Kan PY, Collins LL, Chang C: Proteasome activity is required for androgen receptor transcriptional activity via regulation of androgen receptor nuclear translocation and interaction with coregulators in prostate cancer cells. J Biol Chem. 2002, 277 (39): 36570-36576. 10.1074/jbc.M204751200.View ArticlePubMedGoogle Scholar
- Wu Y, Kawate H, Ohnaka K, Nawata H, Takayanagi R: Nuclear compartmentalization of N-CoR and its interactions with steroid receptors. Mol Cell Biol. 2006, 26 (17): 6633-6655. 10.1128/MCB.01534-05.PubMed CentralView ArticlePubMedGoogle Scholar
- Hsu CL, Chen YL, Ting HJ, Lin WJ, Yang Z, Zhang Y, Wang L, Wu CT, Chang HC, Yeh S: Androgen receptor (AR) NH2- and COOH-terminal interactions result in the differential influences on the AR-mediated transactivation and cell growth. Mol Endocrinol. 2005, 19 (2): 350-361.View ArticlePubMedGoogle Scholar
- Bevan CL, Hoare S, Claessens F, Heery DM, Parker MG: The AF1 and AF2 domains of the androgen receptor interact with distinct regions of SRC1. Mol Cell Biol. 1999, 19 (12): 8383-8392.PubMed CentralView ArticlePubMedGoogle Scholar
- Kaighn ME, Narayan KS, Ohnuki Y, Lechner JF, Jones LW: Establishment and characterization of a human prostatic carcinoma cell line (PC-3). Invest Urol. 1979, 17 (1): 16-23.PubMedGoogle Scholar
- Jensen FC, Girardi AJ, Gilden RV, Koprowski H: Infection of Human and Simian Tissue Cultures with Rous Sarcoma Virus. Proc Natl Acad Sci USA. 1964, 52: 53-59. 10.1073/pnas.52.1.53.PubMed CentralView ArticlePubMedGoogle Scholar
- Rubino D, Driggers P, Arbit D, Kemp L, Miller B, Coso O, Pagliai K, Gray K, Gutkind S, Segars J: Characterization of Brx, a novel Dbl family member that modulates estrogen receptor action. Oncogene. 1998, 16 (19): 2513-2526. 10.1038/sj.onc.1201783.View ArticlePubMedGoogle Scholar
- Butler LM, Centenera MM, Neufing PJ, Buchanan G, Choong CS, Ricciardelli C, Saint K, Lee M, Ochnik A, Yang M: Suppression of androgen receptor signaling in prostate cancer cells by an inhibitory receptor variant. Mol Endocrinol. 2006, 20 (5): 1009-1024. 10.1210/me.2004-0401.View ArticlePubMedGoogle Scholar
- Irvine RA, Ma H, Yu MC, Ross RK, Stallcup MR, Coetzee GA: Inhibition of p160-mediated coactivation with increasing androgen receptor polyglutamine length. Hum Mol Genet. 2000, 9 (2): 267-274. 10.1093/hmg/9.2.267.View ArticlePubMedGoogle Scholar
- Buchanan G, Yang M, Harris JM, Nahm HS, Han G, Moore N, Bentel JM, Matusik RJ, Horsfall DJ, Marshall VR: Mutations at the boundary of the hinge and ligand binding domain of the androgen receptor confer increased transactivation function. Mol Endocrinol. 2001, 15 (1): 46-56. 10.1210/me.15.1.46.View ArticlePubMedGoogle Scholar
- Gordon GW, Berry G, Liang XH, Levine B, Herman B: Quantitative fluorescence resonance energy transfer measurements using fluorescence microscopy. Biophys J. 1998, 74 (5): 2702-2713. 10.1016/S0006-3495(98)77976-7.PubMed CentralView ArticlePubMedGoogle Scholar
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