7b0bfe385c687ef19ad5e796b7de528b pmc_10017050.pdf f328b19081603da571d917446eca69b515378e74 pmc_10017050.pdf 69c1ba7c4f1885df5a1880b732bf995d64b8685ead4d0e596b6eb846d079576d pmc_10017050.pdf Title: A hotspot for posttranslational modifications on the androgen receptor dimer interface drives pathology and anti-androgen resistance Author: Andrea Alegre-Martí, Alba Jiménez-Panizo, Adrián Martínez-Tébar, Coralie Poulard, M. Núria Peralta-Moreno, Montserrat Abella, Rosa Antón, Marcos Chiñas, Ulrich Eckhard, Josep M. Piulats, Ana M. Rojas, Juan Fernández-Recio, Jaime Rubio-Martínez, Muriel Le Romancer, Álvaro Aytes, Pablo Fuentes-Prior, Eva Estébanez-Perpiñá Creator: Servigistics Arbortext Advanced Print Publisher 11.1.4546/W-x64 Producer: PDFlib+PDI 9.0.7p3 (C++/Win64) CreationDate: Fri Mar 10 16:55:24 2023 CET ModDate: Fri Mar 10 16:55:24 2023 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 26 Encrypted: no Page size: 593.972 x 756 pts Page rot: 0 File size: 3964920 bytes Optimized: yes PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SHLQTZ+MyriadPro-Bold Type 1C Custom yes yes yes 432 0 BKEYVB+MyriadPro-Semibold Type 1C Custom yes yes yes 437 0 IBIFVO+SegoeUISymbol CID TrueType Identity-H yes yes yes 442 0 IFMNJJ+STIXGeneral-Bold Type 1C Custom yes yes yes 447 0 GCRXCC+MyriadPro-Regular Type 1C Custom yes yes yes 452 0 KZTRDM+MinionPro-Regular Type 1C Custom yes yes yes 457 0 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CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: OneColumn PageMode: UseNone Language: en Outlines: Item: Title: INTRODUCTION Destination: sec1 Item: Title: RESULTS Destination: sec2 Children: Item: Title: Mutations at the AR-LBD dimer interface differently affect cell proliferation and response to anti-androgens Destination: sec3 Item: Title: Mutations at the AR-LBD dimer interface profoundly deregulate androgen-dependent transcription programs Destination: sec4 Item: Title: AR-LBD interface residue mutations do not compromise domain folding but lead to reduced thermal stability and increased sensitivity to proteolysis Destination: sec5 Item: Title: AR-LBD dimer interface mutants exhibit enhanced flexibility and distinct structural rearrangements both locally and at distant AF-2 and BF-3 sites Destination: sec6 Item: Title: The noncanonical AR-LBD dimerization mode is conserved upon single point mutations of dimer interface residues Destination: sec7 Item: Title: The recurrent point mutation Y764C highlights the ability of AR-LBD dimer interface to adopt different relative orientations in AR homodimers Destination: sec8 Item: Title: The R761 zone undergoes large conformational rearrangements in AR-LBD point mutants Destination: sec9 Item: Title: Full-length AR is preferentially monomethylated in vivo by the methyltransferase, PRMT5, and methylation of Arg761 is affected by AR-LBD dimer interface disease-linked mutations Destination: sec10 Item: Title: DISCUSSION Destination: sec11 Item: Title: MATERIALS AND METHODS Destination: sec12 Children: Item: Title: Cell lines and plasmids Destination: sec13 Item: Title: Luciferase reporter assay Destination: sec14 Item: Title: Functional cell assays Destination: sec15 Children: Item: Title: Colony formation and proliferation assays Destination: sec16 Item: Title: Proliferation assays Destination: sec17 Item: Title: MTT cell viability assays Destination: sec18 Item: Title: Immunoprecipitation and Western blotting Destination: sec19 Item: Title: Transcriptomic analysis Destination: sec20 Item: Title: Enrichment analysis Destination: sec21 Item: Title: AR target genes expression analysis by RT-qPCR Destination: sec22 Item: Title: Proximity ligation assay Destination: sec23 Item: Title: Cloning and mutagenesis of the hAR-LBD construct Destination: sec24 Item: Title: Differential scanning fluorimetry Destination: sec25 Item: Title: Proteolytic digestion Destination: sec26 Item: Title: Crystallization and structure determination Destination: sec27 Item: Title: Cross-linking experiments Destination: sec28 Item: Title: Y764C covalent dimerization in solution Destination: sec29 Item: Title: Nano liquid chromatography MS/MS experiments Destination: sec30 Item: Title: Docking experiments and dimeric interface analysis Destination: sec31 Item: Title: Computational analysis of the estimated energetic impact of AR point mutations Destination: sec32 Item: Title: Statistical coupling analysis Destination: sec33 Item: Title: Molecular dynamic simulations Destination: sec34 Children: Item: Title: System preparation Destination: sec35 Item: Title: Molecular dynamics Destination: sec36 Item: Title: RMSD and RMSF Destination: sec37 Item: Title: Residue pair distance Destination: sec38 Item: Title: Supplementary Materials Destination: sec39 Children: Item: Title: This PDF file includes: Destination: sec40 Item: Title: References and Notes Destination: reflist Item: Title: Acknowledgments Destination: ackBk Info: Title: A hotspot for posttranslational modifications on the androgen receptor dimer interface drives pathology and anti-androgen resistance Author: Andrea Alegre-Martí, Alba Jiménez-Panizo, Adrián Martínez-Tébar, Coralie Poulard, M. Núria Peralta-Moreno, Montserrat Abella, Rosa Antón, Marcos Chiñas, Ulrich Eckhard, Josep M. Piulats, Ana M. 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Núria Peralta-Moreno, Montserrat Abella, Rosa Antón, Marcos Chiñas, Ulrich Eckhard, Josep M. Piulats, Ana M. 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