Karen M. Ong a, John A. Blackford Jr. b, Benjamin L. Kagan b,1, S. Stoney Simons Jr. b,2, and Carson C. Chow a,2
-Author Affiliations
aLaboratory of Biological Modeling, National Institute of Diabetes and Digestive and Kidney Diseases, and
bSteroid Hormones Section, National Institute of Diabetes and Digestive and Kidney Diseases/Clinical Endocrinology Branch, National Institutes of Health, Bethesda, MD 20892
↵1Present address: Department of Oncology, Lombardi Cancer Center, Georgetown University, Washington, DC 20057.
Edited by Keith R. Yamamoto, University of California, San Francisco, CA, and approved March 2, 2010 (received for review September 25, 2009)
Abstract
Ligand-mediated gene induction by steroid receptors is a multistep process characterized by a dose–response curve for gene product that follows a first-order Hill equation. This behavior has classically been explained by steroid binding to receptor being the rate-limiting step. However, this predicts a constant potency of gene induction (EC50) for a given receptor-steroid complex, which is challenged by the findings that various cofactors/reagents can alter this parameter in a gene-specific manner. These properties put strong constraints on the mechanisms of gene induction and raise two questions: How can a first-order Hill dose–response curve (FHDC) arise from a multistep reaction sequence, and how do cofactors modify potency? Here we introduce a theoretical framework in which a sequence of steps yields an FHDC for the final product as a function of the initial agonist concentration. An exact determination of all constants is not required to describe the final FHDC. The theory predicts mechanisms for cofactor/reagent effects on gene-induction potency and maximal activity and it assigns a relative order to cofactors in the sequence of steps. The theory is supported by several observations from glucocorticoid receptor-mediated gene induction. It identifies the mechanism and matches the measured dose–response curves for different concentrations of the combination of cofactor Ubc9 and receptor. It also predicts that an FHDC cannot involve the DNA binding of preformed receptor dimers, which is validated experimentally. The theory is general and can be applied to any biochemical reaction that shows an FHDC.
dose-response Michaelis-Menten gene expression steroid receptors glucocorticoids pharmacology
Footnotes
2To whom correspondence may be addressed. E-mail: carsonc@mail.nih.govor steroids@niddk.nih.gov.
Author contributions: S.S.S. and C.C.C. designed research; K.M.O., J.A.B., B.L.K., S.S.S., and C.C.C. performed research; K.M.O. and C.C.C. performed the mathematical analysis; K.M.O., S.S.S., and C.C.C. analyzed data; and K.M.O., S.S.S., and C.C.C. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/cgi/content/full/0911095107/DCSupplemental.
+++++
