Cooperative DNA Binding by the B-Isoform of Human Progesterone Receptor:  Thermodynamic Analysis Reveals Strongly Favorable and Unfavorable Contributions to Assembly

Heneghan, Aaron F.; Connaghan‐Jones, Keith D.; Miura, Michael T.; Bain, David L.

doi:10.1021/bi052046g

Cited by 24 publications

(52 citation statements)

References 49 publications

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“…These signals originate four to five bases outside the PRE and likely arise because of receptormediated DNA bending (15,16). The more intense hypersensitive signal located equidistant between the two PREs (large arrow) is seen only with the multisite PRE 2 promoter; it localizes to two to three base pairs and has been previously interpreted to be due to cooperative receptor interactions between the response elements (12).…”

Section: Resultsmentioning

confidence: 91%

“…For promoters containing two tandemly linked PREs (PRE 2 ), dimer binding is coupled to cooperative interactions between each site (12,13). This series of reactions is presented schematically in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We present here a thermodynamic dissection of PR-A interactions with a multisite promoter and compare the results to our previous work on PR-B (12). The analysis makes it clear that whereas PR-B and PR-A appear to have similar binding affinities, the actual energetics of assembly are considerably different, both at the level of the intrinsic binding affinity and the extent of cooperative interactions between palindromic PREs.…”

mentioning

confidence: 78%

“…Our earlier work on the energetics of PR-B selfassembly and cooperative DNA binding found that liganded, preformed solution dimers interact with individual response elements with picomolar, rather than nanomolar binding affinity (11,12). Moreover, binding to multiple response elements is coupled to highly cooperative assembly reactions.…”

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confidence: 99%

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Thermodynamic analysis of progesterone receptor–promoter interactions reveals a molecular model for isoform-specific function

Connaghan‐Jones

Heneghan²,

Miura³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Human progesterone receptors (PR) exist as two functionally distinct isoforms, PR-A and PR-B. The proteins are identical except for an additional 164 residues located at the N terminus of PR-B. To determine the mechanisms responsible for isoform-specific functional differences, we present here a thermodynamic dissection of PR-A-promoter interactions and compare the results to our previous work on PR-B. This analysis has generated a number of results inconsistent with the traditional, biochemically based model of receptor function. Specifically, statistical models invoking preformed PR-A dimers as the active binding species demonstrate that intrinsic binding energetics are over an order of magnitude greater than is apparent. High-affinity binding is opposed, however, by a large energetic penalty. The consequences of this penalty are 2-fold: Successive monomer binding to a palindromic response element is thermodynamically favored over preformed dimer binding, and DNA-induced dimerization of the monomers is largely abolished. Furthermore, PR-A binding to multiple PREs is only weakly cooperative, as judged by a 5-fold increase in overall stability. Comparison of these results to our work on PR-B demonstrates that whereas both isoforms appear to have similar DNA binding affinities, PR-B in fact has a greatly increased intrinsic binding affinity and cooperative binding ability relative to PR-A. These differences thus suggest that residues unique to PR-B allosterically regulate the energetics of cooperative promoter assembly. From a functional perspective, the differences in microscopic affinities predict receptor-promoter occupancies that accurately correlate with the transcriptional activation profiles seen for each isoform.nuclear receptor ͉ protein-DNA interactions ͉ quantitative footprint titrations ͉ statistical mechanics

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Section: Resultsmentioning

confidence: 91%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 78%

mentioning

confidence: 99%

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Thermodynamic analysis of progesterone receptor–promoter interactions reveals a molecular model for isoform-specific function

Connaghan‐Jones

Heneghan²,

Miura³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…Proposed mechanisms include cooperative recruitment of coactivators, action at distinct rate-limiting steps in initiation, cooperative DNA binding, and/or direct protein-protein interactions between receptor dimers (56,76). It is possible that the strong transcriptional activity of PR-B on tandem PREs is due to cooperative interactions between receptor dimers bound at adjacent PREs rather than between monomers bound at a single PRE.…”

Section: Discussionmentioning

confidence: 99%

Mechanisms Underlying the Control of Progesterone Receptor Transcriptional Activity by SUMOylation

Abdel-Hafiz

Dudevoir

Horwitz

2009

Journal of Biological Chemistry

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Posttranslational modification by small ubiquitin-like modifier (SUMO) is a major regulator of transcription. We previously showed that progesterone receptors (PR) have a single consensus KXE SUMO-conjugation motif centered at Lys-388 in the N-terminal domain of PR-B and a homologous site of PR-A. SUMOylation of the PR is hormone-dependent and has a suppressive effect on transcription of an exogenous promoter. Here we show that repression of PR activity by SUMOylation at Lys-388 is uncoupled from phosphorylation, involves synergy between tandem progesterone response elements, and is associated with lowered ligand sensitivity and slowed ligand-dependent down-regulation. However, paradoxically, cellular overexpression of SUMO-1 increases PR transcriptional activity even if Lys-388 is mutated, suggesting that the receptors are activated indirectly by other SUMOylated proteins. One of these is the coactivator SRC-1, whose binding to PR and enhancement of agonist-dependent N-/C-terminal interactions is augmented by the presence of SUMO-1. Increased transcription due to SRC-1 is independent of PR SUMOylation based on assays with the Lys-388 mutants and the pure antiprogestin ZK98299, which blocks N-/C-terminal interactions. In summary, SUMOylation tightly regulates the transcriptional activity of PR by repressing the receptors directly while activating them indirectly through augmented SRC-1 coactivation.

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Steroid Hormones

Simons¹

2008

Wiley Encyclopedia of Chemical Biology

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Five categories of steroid hormones exist in humans, including androgens, estrogens, glucocorticoids, mineralocorticoids, and progestins. These hormones affect virtually every tissue and organ in the human body and play major roles in the development, differentiation, and homeostasis of normal individuals. Antisteroids usually possess nonsteroidal structures but still block the actions of the steroid hormones and are important tools in endocrine therapies of pathologic conditions. Therefore, how the body regulates where, when, and how much a response to steroids occurs is of major importance. Here we survey what is known about the genomic responses to steroid hormones, each of which is mediated by a unique intracellular receptor protein that interacts with the cellular DNA to modify the rates of gene transcription. These receptors are members of a much larger superfamily of steroid/nuclear receptors, most of which bind either nonsteroidal ligands or no known ligand. Nongenomic (i.e., pathways without initial involvement of genomic DNA) and secondary responses (i.e., changes that require protein synthesis to alter gene transcription) are additional important effects of steroid hormones but are not discussed here. The emphasis is on the biochemistry of the five classes of steroid hormones, the techniques used to study steroid hormone action, and the basic mechanistic steps by which steroids alter gene expression.

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Cooperative DNA Binding by the B-Isoform of Human Progesterone Receptor: Thermodynamic Analysis Reveals Strongly Favorable and Unfavorable Contributions to Assembly

Cited by 24 publications

References 49 publications

Thermodynamic analysis of progesterone receptor–promoter interactions reveals a molecular model for isoform-specific function

Thermodynamic analysis of progesterone receptor–promoter interactions reveals a molecular model for isoform-specific function

Mechanisms Underlying the Control of Progesterone Receptor Transcriptional Activity by SUMOylation

Steroid Hormones

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