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Molecular Mechanisms of Androgen Receptor Functions

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Molecular Mechanisms of Androgen Receptor Functions Molecular Mechanisms of Androgen Receptor Functions Moleculaire mechanismen van androgeenreceptor functies Proefschrift ter verkrijging van de graad van doctor aan de Erasmus Universiteit Rotterdam op gezag van de rector magnificus Prof.dr. S.W.J. Lamberts en volgens besluit van het College voor Promoties De openbare verdediging zal plaatsvinden op woensdag 21 november 2007 om 9.45 uur door Karine Steketee geboren te 's-Heer Arendskerke Promotiecommissie Promotor : Prof.dr.ir. J. Trapman Overige leden : Prof.dr. J.A. Grootegoed Prof.dr. J.P.T.M. van Leeuwen Dr.ir. G.W. Jenster Copromotor: Dr. A.O. Brinkmann The work described in this thesis was performed at the Department of Pathology of the Josephine Nefkens Institute, Erasmus MC, Rotterdam, the Netherlands. Printed by: Wöhrmann, Zutphen ISBN: 978-90-8570-295-5 2 In dankbare herinnering aan mijn vader Voor mijn moeder Voor Jeroen 3 Contents List of abbreviations.....................................................................................................................6 Chapter 1 General introduction.................................................................................................9 INTRODUCTION ................................................................................................................................................... 10 1.1 NUCLEAR RECEPTORS.............................................................................................................................. 10 1.1.1 General features of nuclear receptors ..................................................................................................... 10 1.1.2 Structure and functional domains of nuclear receptors........................................................................... 11 1.1.2.1 The amino-terminal domain.............................................................................................................. 11 1.1.2.2 The DNA binding domain ................................................................................................................ 12 1.1.2.3 The hinge region............................................................................................................................... 12 1.1.2.4 The ligand binding domain............................................................................................................... 13 1.1.3 The nuclear receptor subfamily of steroid receptors............................................................................... 14 1.1.4 Nuclear receptors in transcription regulatory complexes ....................................................................... 15 1.1.5 Nuclear receptor coactivators ................................................................................................................. 18 1.1.6 Nuclear receptor corepressors ................................................................................................................ 20 1.1.7 Post-translational modifications of nuclear receptors ............................................................................ 21 1.1.8 Additional nuclear receptor functions ..................................................................................................... 22 1.2 THE ANDROGEN RECEPTOR.................................................................................................................... 24 1.2.1 General features of the AR ...................................................................................................................... 24 1.2.1.1 The AR NH 2-terminal domain.......................................................................................................... 25 1.2.1.2 The AR DNA binding domain.......................................................................................................... 25 1.2.1.3 The AR hinge region ........................................................................................................................ 26 1.2.1.4 The AR ligand binding domain......................................................................................................... 27 1.2.2 AR coregulators....................................................................................................................................... 28 1.2.2.1 AR coactivators ................................................................................................................................ 29 1.2.2.2 AR corepressors................................................................................................................................ 33 1.2.2.3 The AR in transcription regulatory complexes ................................................................................. 35 1.2.2.4 Post-translational modifications of the AR........................................................... 37 1.2.3 AR LBD interaction with AR NTD and AR coactivators.......................................................................... 39 1.2.3.1 AR N/C interaction........................................................................................................................... 39 1.2.3.2 Relevance of AR N/C interaction in receptor function..................................................................... 40 1.2.3.3 N/C interaction in other nuclear receptors ........................................................................................ 41 1.2.3.4 AR LBD specifically binds to FXXLF motifs in AR NTD and AR coactivators............................. 42 1.2.3.4.1 Structural background of the preference for FXXLF motifs of the AR coactivator groove ....... 42 1.2.3.4.2 Variations of the FXXLF motif ................................................................................................. 44 1.2.4 Androgen-specific gene transcription...................................................................................................... 45 1.2.4.1 General mechanism of androgen regulated gene transcription ......................................................... 45 1.2.4.2 Androgen-specifically regulated genes............................................................................................. 45 1.2.4.3 Mechanism of AR binding to androgen-specific AREs.................................................................... 48 1.2.4.4 Role of androgen-specific AREs ...................................................................................................... 49 1.2.5 The AR in disease .................................................................................................................................... 51 1.2.5.1 The AR in prostate cancer ................................................................................................................ 51 1.2.5.1.1 Prostate cancer development and progression ........................................................................... 51 1.2.5.1.2 Androgen-dependent prostate cancer.......................................................................................... 52 1.2.5.1.3 Mechanisms of AR activation in androgen-independent prostate cancer ....................................... 52 1.2.5.1.3.1 AR amplification and overexpression................................................................................. 53 1.2.5.1.3.2 AR mutations ...................................................................................................................... 53 1.2.5.1.3.3 Intraprostatic conversion of adrenal androgens to T and DHT............................................... 56 1.2.5.1.3.4 Ligand-independent activation of the AR through cross-talk with other signaling pathways 57 1.2.5.1.3.5 Aberrant AR coregulator expression and function.................................................................. 58 1.2.5.1.3.6 Signaling pathways in prostate cancer cells bypassing the AR............................................... 59 1.3 AIM AND SCOPE OF THIS THESIS........................................................................................................... 60 1.4 REFERENCES ............................................................................................................................................... 61 4 Chapter 2 Amino acids 3-13 and amino acids in and flanking the 23 FXXLF 27 motif modulate the interaction between the amino-terminal and ligand-binding domain of the androgen receptor ......................................................................................................................89 Chapter 3 A bioinformatics-based functional analysis shows that the specifically androgen-regulated gene SARG contains an active direct repeat androgen response element in the first intron. .......................................................................................................109 Chapter 4 Broadened ligand responsiveness of androgen receptor mutants obtained by random amino acid substitution of H874 and mutation hot spot T877 in prostate cancer ....................................................................................................................................................131 Chapter 5 General discussion and future perspectives........................................................149 5.1 A IM OF AR RESEARCH ................................................................................................................................... 150 5.2 M OLECULAR MECHANISMS OF AR FUNCTIONS............................................................................................... 150 5.2.1 N/C interaction .....................................................................................................................................
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