Influence of Domain Interactions on Conformational Mobility of the Progesterone Receptor Detected by Hydrogen/Deuterium Exchange Mass Spectrometry

• Published in: Structure (London) 2014-07, Vol.22 (7), p.961-973
• Main Authors: Goswami, Devrishi, Callaway, Celetta, Pascal, Bruce D., Kumar, Raj, Edwards, Dean P., Griffin, Patrick R.
• Format: Article
• Language: English
• Subjects: Activation; Activation analysis; Amino Acid Sequence; Animals; Deuterium; Deuterium Exchange Measurement; Dominance; Exchange; Humans; Ligands; Mass spectrometry; Mass Spectrometry - methods; Mathematical models; Mifepristone - chemistry; Mifepristone - metabolism; Models, Molecular; Molecular Sequence Data; Promegestone - chemistry; Promegestone - metabolism; Protein Binding; Protein Conformation; Protein Structure, Tertiary; Receptors; Receptors, Cytoplasmic and Nuclear - chemistry; Receptors, Cytoplasmic and Nuclear - metabolism; Receptors, Progesterone - chemistry; Receptors, Progesterone - genetics; Receptors, Progesterone - metabolism; Sf9 Cells; TATA-Box Binding Protein - chemistry; TATA-Box Binding Protein - metabolism
• ISSN: 0969-2126; 1878-4186
• DOI: 10.1016/j.str.2014.04.013

Structural and functional details of the N-terminal activation function 1 (AF1) of most nuclear receptors are poorly understood due to the highly dynamic intrinsically disordered nature of this domain. A hydrogen/deuterium exchange (HDX) mass-spectrometry-based investigation of TATA box-binding protein (TBP) interaction with various domains of progesterone receptor (PR) demonstrate that agonist-bound PR interaction with TBP via AF1 impacts the mobility of the C-terminal AF2. Results from HDX and other biophysical studies involving agonist- and antagonist-bound full-length PR and isolated PR domains reveal the molecular mechanism underlying synergistic transcriptional activation mediated by AF1 and AF2, dominance of PR-B isoform over PR-A, and the necessity of AF2 for full AF1-mediated transcriptional activity. These results provide a comprehensive picture elaborating the underlying mechanism of PR-TBP interactions as a model for studying nuclear receptor (NR)-transcription factor functional interactions. [Display omitted] •Interdomain communication with PR upon coactivator binding detected by HDX•Molecular basis of AF1-AF2 synergism for TBP-dependent PR activation•Conformational mobility of full-length PR isoforms (∼100 kDa) characterized by HDX•Isoform-specific interactions detected by HDX Goswami et al. present a hydrogen/deuterium exchange investigation of TATA box binding protein (TBP) interaction with the progesterone receptor (PR). Agonist-bound PR interaction with TBP via AF1 impacts the mobility of the C-terminal AF2, underscoring synergistic, AF1 and AF2 mediated, transcriptional activation.