Site-Specific Phosphorylation Induces Functionally Active Conformation in the Intrinsically Disordered N-Terminal Activation Function (AF1) Domain of the Glucocorticoid Receptor

Abstract

Intrinsically disordered (ID) regions are disproportionately higher in cell signaling proteins and are predicted to have much larger frequency of phosphorylation sites than ordered regions, suggesting an important role in their regulatory capacity. In this study, we show that AF1, an ID activation domain of the glucocorticoid receptor (GR), adopts a functionally folded conformation due to its site-specific phosphorylation by p38 mitogen-activated protein kinase, which is involved in apoptotic and gene-inductive events initiated by the GR. Further, we show that site-specific phosphorylation-induced secondary and tertiary structure formation specifically facilitates AF1's interaction with critical coregulatory proteins and subsequently its transcriptional activity. These data demonstrate a mechanism through which ID activation domain of the steroid receptors and other similar transcription factors may adopt a functionally active conformation under physiological conditions.

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Articles of Significant Interest Selected from This Issue by the Editors

This study was supported by National Institutes of Health grant NIDDK DK058829 (to R.K.).

We thank A. Mishra for technical help in creating some of the AF1 mutants, E. B. Thompson (University of Texas Medical Branch [UTMB]) for providing GR plasmids, D. W. Bolen (UTMB) for CD analysis, M. Mancini (Baylor College of Medicine) for the pYFP-CBP and pYFP-SRC-1 constructs, and L. Vergara and the Optical Imaging Core staff (UTMB) for technical support and help with data analysis.