Structural basis for the regulation of β-glucuronidase expression by human gut Enterobacteriaceae

Little, Michael S.; Pellock, Samuel J.; Walton, William G.; Tripathy, Ashutosh; Redinbo, Matthew R.

doi:10.1073/pnas.1716241115

Structural basis for the regulation of β-glucuronidase expression by human gut Enterobacteriaceae

Journal Article · Wed Dec 20 23:00:00 EST 2017 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1716241115· OSTI ID:1462565

Little, Michael S. ^[1]; Pellock, Samuel J. ^[1]; Walton, William G. ^[1]; Tripathy, Ashutosh ^[1]; Redinbo, Matthew R. ^[1]

Univ. of North Carolina, Chapel Hill, NC (United States)

The gut microbiota harbor diverse β-glucuronidase (GUS) enzymes that liberate glucuronic acid (GlcA) sugars from small-molecule conjugates and complex carbohydrates. However, only the Enterobacteriaceae family of human gut-associated Proteobacteria maintain a GUS operon under the transcriptional control of a glucuronide repressor, GusR. Despite its potential importance in Escherichia, Salmonella, Klebsiella, Shigella, and Yersinia opportunistic pathogens, the structure of GusR has not been examined. Here, we explore the molecular basis for GusR-mediated regulation of GUS expression in response to small-molecule glucuronides. Presented are 2.1-Å-resolution crystal structures of GusRs from Escherichia coli and Salmonella enterica in complexes with a glucuronide ligand. The GusR-specific DNA operator site in the regulatory region of the E. coli GUS operon is identified, and structure-guided GusR mutants pinpoint the residues essential for DNA binding and glucuronide recognition. Interestingly, the endobiotic estradiol-17-glucuronide and the xenobiotic indomethacin-acyl-glucuronide are found to exhibit markedly differential binding to these GusR orthologs. Using structure-guided mutations, we are able to transfer E. coli GusR’s preferential DNA and glucuronide binding affinity to S. enterica GusR. Structures of putative GusR orthologs from GUS-encoding Firmicutes species also reveal functionally unique features of the Enterobacteriaceae GusRs. Finally, dominant-negative GusR variants are validated in cell-based studies. In conclusion, these data provide a molecular framework toward understanding the control of glucuronide utilization by opportunistic pathogens in the human gut.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

OSTI ID:: 1462565

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 2 Vol. 115; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
biochemistry
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molecular biology
structural biology
transcriptional regulation

Structural basis for the regulation of β-glucuronidase expression by human gut Enterobacteriaceae

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