skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Structure and Function of the Macrolide Biosensor Protein, MphR(A), with and without Erythromycin

Abstract

The regulatory protein MphR(A) has recently seen extensive use in synthetic biological applications, such as metabolite sensing and exogenous control of gene expression. This protein negatively regulates the expression of a macrolide 2{prime}-phosphotransferase I resistance gene (mphA) via binding to a 35-bp DNA operator upstream of the start codon and is de-repressed by the presence of erythromycin. Here, we present the refined crystal structure of the MphR(A) protein free of erythromycin and that of the MphR(A) protein with bound erythromycin at 2.00- and 1.76-{angstrom} resolutions, respectively. We also studied the DNA binding properties of the protein and identified mutants of MphR(A) that are defective in gene repression and ligand binding in a cell-based reporter assay. The combination of these two structures illustrates the molecular basis of erythromycin-induced gene expression and provides a framework for additional applied uses of this protein in the isolation and engineered biosynthesis of polyketide natural products.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1005783
Resource Type:
Journal Article
Journal Name:
J. Mol. Biol.
Additional Journal Information:
Journal Volume: 387; Journal Issue: (5) ; 04, 2009; Journal ID: ISSN 0022-2836
Country of Publication:
United States
Language:
ENGLISH
Subject:
36 MATERIALS SCIENCE; BIOSYNTHESIS; CODONS; CRYSTAL STRUCTURE; DNA; ERYTHROMYCIN; GENES; METABOLITES; MUTANTS; PROTEINS

Citation Formats

Zheng, Jianting, Sagar, Vatsala, Smolinsky, Adam, Bourke, Chase, LaRonde-LeBlanc, Nicole, Cropp, T Ashton, and Maryland). Structure and Function of the Macrolide Biosensor Protein, MphR(A), with and without Erythromycin. United States: N. p., 2009. Web. doi:10.1016/j.jmb.2009.02.058.
Zheng, Jianting, Sagar, Vatsala, Smolinsky, Adam, Bourke, Chase, LaRonde-LeBlanc, Nicole, Cropp, T Ashton, & Maryland). Structure and Function of the Macrolide Biosensor Protein, MphR(A), with and without Erythromycin. United States. https://doi.org/10.1016/j.jmb.2009.02.058
Zheng, Jianting, Sagar, Vatsala, Smolinsky, Adam, Bourke, Chase, LaRonde-LeBlanc, Nicole, Cropp, T Ashton, and Maryland). 2009. "Structure and Function of the Macrolide Biosensor Protein, MphR(A), with and without Erythromycin". United States. https://doi.org/10.1016/j.jmb.2009.02.058.
@article{osti_1005783,
title = {Structure and Function of the Macrolide Biosensor Protein, MphR(A), with and without Erythromycin},
author = {Zheng, Jianting and Sagar, Vatsala and Smolinsky, Adam and Bourke, Chase and LaRonde-LeBlanc, Nicole and Cropp, T Ashton and Maryland)},
abstractNote = {The regulatory protein MphR(A) has recently seen extensive use in synthetic biological applications, such as metabolite sensing and exogenous control of gene expression. This protein negatively regulates the expression of a macrolide 2{prime}-phosphotransferase I resistance gene (mphA) via binding to a 35-bp DNA operator upstream of the start codon and is de-repressed by the presence of erythromycin. Here, we present the refined crystal structure of the MphR(A) protein free of erythromycin and that of the MphR(A) protein with bound erythromycin at 2.00- and 1.76-{angstrom} resolutions, respectively. We also studied the DNA binding properties of the protein and identified mutants of MphR(A) that are defective in gene repression and ligand binding in a cell-based reporter assay. The combination of these two structures illustrates the molecular basis of erythromycin-induced gene expression and provides a framework for additional applied uses of this protein in the isolation and engineered biosynthesis of polyketide natural products.},
doi = {10.1016/j.jmb.2009.02.058},
url = {https://www.osti.gov/biblio/1005783}, journal = {J. Mol. Biol.},
issn = {0022-2836},
number = (5) ; 04, 2009,
volume = 387,
place = {United States},
year = {Wed Sep 02 00:00:00 EDT 2009},
month = {Wed Sep 02 00:00:00 EDT 2009}
}