title: Regulation of the structurally dynamic N-terminal domain of progesterone receptor by protein-induced folding.
abstract: The N-terminal domain (NTD) of steroid receptors harbors a transcriptional activation function (AF1) that is composed of an intrinsically disordered polypeptide. We examined the interaction of the TATA-binding protein (TBP) with the NTD of the progesterone receptor (PR) and its ability to regulate AF1 activity through coupled folding and binding. As assessed by solution phase biophysical methods, the isolated NTD of PR contains a large content of random coil, and it is capable of adopting secondary α-helical structure and more stable tertiary folding either in the presence of the natural osmolyte trimethylamine-N-oxide or through a direct interaction with TBP. Hydrogen-deuterium exchange coupled with mass spectrometry confirmed the highly dynamic intrinsically disordered property of the NTD within the context of full-length PR. Deletion mapping and point mutagenesis defined a region of the NTD (amino acids 350-428) required for structural folding in response to TBP interaction. Overexpression of TBP in cells enhanced transcriptional activity mediated by the PR NTD, and deletion mutations showed that a region (amino acids 327-428), similar to that required for TBP-induced folding, was required for functional response. TBP also increased steroid receptor co-activator 1 (SRC-1) interaction with the PR NTD and cooperated with SRC-1 to stimulate NTD-dependent transcriptional activity. These data suggest that TBP can mediate structural reorganization of the NTD to facilitate the binding of co-activators required for maximal transcriptional activation.
authors.affiliation: the Department of Basic Sciences, Commonwealth Medical College, Scranton, Pennsylvania 18509.
authors.forename: Raj
authors.initials: R
authors.lastname: Kumar
authors.affiliation: From the Departments of Molecular and Cellular Biology and.
authors.forename: Carmen M
authors.initials: CM
authors.lastname: Moure
authors.affiliation: the Department of Basic Sciences, Commonwealth Medical College, Scranton, Pennsylvania 18509.
authors.forename: Shagufta H
authors.initials: SH
authors.lastname: Khan
authors.affiliation: From the Departments of Molecular and Cellular Biology and.
authors.forename: Celetta
authors.initials: C
authors.lastname: Callaway
authors.affiliation: From the Departments of Molecular and Cellular Biology and.
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Grimm
authors.affiliation: the Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida 33458, and.
authors.forename: Devrishi
authors.initials: D
authors.lastname: Goswami
authors.affiliation: the Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida 33458, and.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Griffin
authors.affiliation: From the Departments of Molecular and Cellular Biology and; Pathology and Immunology, Baylor College of Medicine, Houston, Texas 77030,. Electronic address: deane@bcm.edu.
authors.forename: Dean P
authors.initials: DP
authors.lastname: Edwards
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006801:Humans; D056920:Nuclear Receptor Coactivator 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017354:Point Mutation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011980:Receptors, Progesterone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D035181:TATA-Box Binding Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation / Q000502:physiology*
chemical_list: D011980:Receptors, Progesterone; D035181:TATA-Box Binding Protein; C509817:TBP protein, human; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1
keywords: Hydrogen Deuterium Exchange Mass; Intrinsically Disordered Proteins; Mass Spectrometry (MS); N-terminal Transcription Activation domain AF1; Progesterone; Progesterone Receptor; Protein Folding; Steroid Hormone Receptor; TATA-binding Protein