Matches in SemOpenAlex for { <https://semopenalex.org/work/W2333259448> ?p ?o ?g. }
- W2333259448 endingPage "418" @default.
- W2333259448 startingPage "395" @default.
- W2333259448 abstract "The androgen receptor–transcriptional intermediary factor 2 (AR-TIF2) positional protein–protein interaction (PPI) biosensor assay described herein combines physiologically relevant cell-based assays with the specificity of binding assays by incorporating structural information of AR and TIF2 functional domains along with intracellular targeting sequences and fluorescent reporters. Expression of the AR-red fluorescent protein (RFP) “prey” and TIF2-green fluorescent protein (GFP) “bait” components of the biosensor was directed by recombinant adenovirus constructs that expressed the ligand binding and activation function 2 surface domains of AR fused to RFP with nuclear localization and nuclear export sequences, and three α-helical LXXLL motifs from TIF2 fused to GFP and an HIV Rev nucleolar targeting sequence. In unstimulated cells, AR-RFP was localized predominantly to the cytoplasm and TIF2-GFP was localized to nucleoli. Dihydrotestosterone (DHT) treatment induced AR-RFP translocation into the nucleus where the PPIs between AR and TIF2 resulted in the colocalization of both biosensors within the nucleolus. We adapted the translocation enhanced image analysis module to quantify the colocalization of the AR-RFP and TIF2-GFP biosensors in images acquired on the ImageXpress platform. DHT induced a concentration-dependent AR-TIF2 colocalization and produced a characteristic condensed punctate AR-RFP PPI nucleolar distribution pattern. The heat-shock protein 90 inhibitor 17-N-allylamino-17-demethoxygeldanamycin (17-AAG) and antiandrogens flutamide and bicalutamide inhibited DHT-induced AR-TIF2 PPI formation with 50% inhibition concentrations (IC50s) of 88.5±12.5 nM, 7.6±2.4 μM, and 1.6±0.4 μM, respectively. Images of the AR-RFP distribution phenotype allowed us to distinguish between 17-AAG and flutamide, which prevented AR translocation, and bicalutamide, which blocked AR-TIF2 PPIs. We screened the Library of Pharmacologically Active Compounds (LOPAC) set for compounds that inhibited AR-TIF2 PPI formation or disrupted preexisting complexes. Eleven modulators of steroid family nuclear receptors (NRs) and 6 non-NR ligands inhibited AR-TIF2 PPI formation, and 10 disrupted preexisting complexes. The hits appear to be either AR antagonists or nonspecific inhibitors of NR activation and trafficking. Given that the LOPAC set represents such a small and restricted biological and chemical diversity, it is anticipated that screening a much larger and more diverse compound library will be required to find AR-TIF2 PPI inhibitors/disruptors. The AR-TIF2 protein–protein interaction biosensor (PPIB) approach offers significant promise for identifying molecules with potential to modulate AR transcriptional activity in a cell-specific manner that is distinct from the existing antiandrogen drugs that target AR binding or production. Small molecules that disrupt AR signaling at the level of AR-TIF2 PPIs may also overcome the development of resistance and progression to castration-resistant prostate cancer." @default.
- W2333259448 created "2016-06-24" @default.
- W2333259448 creator A5029471010 @default.
- W2333259448 creator A5066405052 @default.
- W2333259448 creator A5076801265 @default.
- W2333259448 creator A5087599491 @default.
- W2333259448 date "2014-09-01" @default.
- W2333259448 modified "2023-09-30" @default.
- W2333259448 title "High-Content Positional Biosensor Screening Assay for Compounds to Prevent or Disrupt Androgen Receptor and Transcriptional Intermediary Factor 2 Protein–Protein Interactions" @default.
- W2333259448 cites W1480912559 @default.
- W2333259448 cites W1544635645 @default.
- W2333259448 cites W1595570100 @default.
- W2333259448 cites W1601895281 @default.
- W2333259448 cites W1965006392 @default.
- W2333259448 cites W1969069329 @default.
- W2333259448 cites W1972158913 @default.
- W2333259448 cites W1979801990 @default.
- W2333259448 cites W1981824495 @default.
- W2333259448 cites W1983203301 @default.
- W2333259448 cites W1983806503 @default.
- W2333259448 cites W1989854777 @default.
- W2333259448 cites W1990529844 @default.
- W2333259448 cites W1997514006 @default.
- W2333259448 cites W2004109773 @default.
- W2333259448 cites W2007172433 @default.
- W2333259448 cites W2008984675 @default.
- W2333259448 cites W2010246016 @default.
- W2333259448 cites W2010583609 @default.
- W2333259448 cites W2017977369 @default.
- W2333259448 cites W2018704721 @default.
- W2333259448 cites W2020195882 @default.
- W2333259448 cites W2022288928 @default.
- W2333259448 cites W2024377156 @default.
- W2333259448 cites W2027181664 @default.
- W2333259448 cites W2027704988 @default.
- W2333259448 cites W2029935944 @default.
- W2333259448 cites W2032363243 @default.
- W2333259448 cites W2033281378 @default.
- W2333259448 cites W2038178452 @default.
- W2333259448 cites W2038954423 @default.
- W2333259448 cites W2040607554 @default.
- W2333259448 cites W2044817493 @default.
- W2333259448 cites W2045166000 @default.
- W2333259448 cites W2048869439 @default.
- W2333259448 cites W2052417817 @default.
- W2333259448 cites W2056169996 @default.
- W2333259448 cites W2059170306 @default.
- W2333259448 cites W2067640792 @default.
- W2333259448 cites W2071857822 @default.
- W2333259448 cites W2072677374 @default.
- W2333259448 cites W2074369446 @default.
- W2333259448 cites W2079039801 @default.
- W2333259448 cites W2079360548 @default.
- W2333259448 cites W2081963757 @default.
- W2333259448 cites W2082163470 @default.
- W2333259448 cites W2085124463 @default.
- W2333259448 cites W2088032617 @default.
- W2333259448 cites W2089267976 @default.
- W2333259448 cites W2089966673 @default.
- W2333259448 cites W2090041442 @default.
- W2333259448 cites W2090796236 @default.
- W2333259448 cites W2092318101 @default.
- W2333259448 cites W2096594220 @default.
- W2333259448 cites W2106323895 @default.
- W2333259448 cites W2107850854 @default.
- W2333259448 cites W2108486460 @default.
- W2333259448 cites W2119504728 @default.
- W2333259448 cites W2134170962 @default.
- W2333259448 cites W2140038897 @default.
- W2333259448 cites W2141255350 @default.
- W2333259448 cites W2143920112 @default.
- W2333259448 cites W2144754627 @default.
- W2333259448 cites W2145777186 @default.
- W2333259448 cites W2151721001 @default.
- W2333259448 cites W2152689753 @default.
- W2333259448 cites W2160314486 @default.
- W2333259448 cites W2161286968 @default.
- W2333259448 cites W2161685546 @default.
- W2333259448 cites W2406991861 @default.
- W2333259448 cites W4232866872 @default.
- W2333259448 cites W4252110563 @default.
- W2333259448 doi "https://doi.org/10.1089/adt.2014.594" @default.
- W2333259448 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4146499" @default.
- W2333259448 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/25181412" @default.
- W2333259448 hasPublicationYear "2014" @default.
- W2333259448 type Work @default.
- W2333259448 sameAs 2333259448 @default.
- W2333259448 citedByCount "18" @default.
- W2333259448 countsByYear W23332594482016 @default.
- W2333259448 countsByYear W23332594482017 @default.
- W2333259448 countsByYear W23332594482018 @default.
- W2333259448 countsByYear W23332594482019 @default.
- W2333259448 countsByYear W23332594482020 @default.
- W2333259448 countsByYear W23332594482021 @default.
- W2333259448 countsByYear W23332594482023 @default.
- W2333259448 crossrefType "journal-article" @default.
- W2333259448 hasAuthorship W2333259448A5029471010 @default.
- W2333259448 hasAuthorship W2333259448A5066405052 @default.