title: The N-terminal domain of the androgen receptor drives its nuclear localization in castration-resistant prostate cancer cells.
abstract: Androgen-independent nuclear localization is required for androgen receptor (AR) transactivation in castration-resistant prostate cancer (CRPC) and should be a key step leading to castration resistance. However, mechanism(s) leading to androgen-independent AR nuclear localization are poorly understood. Since the N-terminal domain (NTD) of AR plays a role in transactivation under androgen-depleted conditions, we investigated the role of the NTD in AR nuclear localization in CRPC. Deletion mutagenesis was used to identify amino acid sequences in the NTD essential for its androgen-independent nuclear localization in C4-2, a widely used CRPC cell line. Deletion mutants of AR tagged with green fluorescent protein (GFP) at the 5'-end were generated and their signal distribution was investigated in C4-2 cells by fluorescent microscopy. Our results showed that the region of a.a. 294-556 was required for androgen-independent AR nuclear localization whereas a.a. 1-293 mediates Hsp90 regulation of AR nuclear localization in CRPC cells. Although the region of a.a. 294-556 does not contain a nuclear import signal, it was able to enhance DHT-induced import of the ligand binding domain (LBD). Also, transactivation of the NTD could be uncoupled from its modulation of AR nuclear localization in C4-2 cells. These observations suggest an important role of the NTD in AR intracellular trafficking and androgen-independent AR nuclear localization in CRPC cells.
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States; Central Laboratory College of Science, King Saud University, Riyadh KSA-11451, Saudi Arabia.
authors.forename: Javid A
authors.initials: JA
authors.lastname: Dar
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States.
authors.forename: Khalid Z
authors.initials: KZ
authors.lastname: Masoodi
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States.
authors.forename: Kurtis
authors.initials: K
authors.lastname: Eisermann
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States.
authors.forename: Sudhir
authors.initials: S
authors.lastname: Isharwal
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States.
authors.forename: Junkui
authors.initials: J
authors.lastname: Ai
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Pascal
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States; University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States.
authors.forename: Joel B
authors.initials: JB
authors.lastname: Nelson
authors.affiliation: Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States; Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15260, United States; University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA 15232, United States. Electronic address: wangz2@upmc.edu.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Wang
mesh_terms: D021581:Active Transport, Cell Nucleus; D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D051958:Nuclear Receptor Coactivator 2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D064129:Prostatic Neoplasms, Castration-Resistant / Q000378:metabolism* / Q000473:pathology; D034741:RNA, Small Interfering / Q000235:genetics; D011944:Receptors, Androgen / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C410798:AR protein, human; D000728:Androgens; D018841:HSP90 Heat-Shock Proteins; C498001:NCOA2 protein, human; D051958:Nuclear Receptor Coactivator 2; D034741:RNA, Small Interfering; D011944:Receptors, Androgen