TY - JOUR
T1 - Cell-based screen for discovering lipopolysaccharide biogenesis inhibitors
AU - Zhang, Ge
AU - Baidin, Vadim
AU - Pahil, Karanbir S.
AU - Moison, Eileen
AU - Tomasek, David
AU - Ramadoss, Nitya S.
AU - Chatterjee, Arnab K.
AU - McNamara, Case W.
AU - Young, Travis S.
AU - Schultz, Peter G.
AU - Meredith, Timothy C.
AU - Kahne, Daniel
N1 - Funding Information:
ACKNOWLEDGMENTS. We thank ICCB-Longwood Screening Facility for providing access to their equipment, the Bauer Core Facility at Harvard University for the use of equipment and sequencing services, and Claudio Zambaldo (The Scripps Research Institute) for providing reagents. We also thank Maria Ericsson, Louise Trakimas, and Elizabeth Benecchi at the Harvard Medical School Electron Microscopy Facility for providing transmission electron microscopy services and for helpful discussions. This work was supported by NIH Grants U19 AI109764 and R01 AI081059 (to D.K.) and the Blavatnik Biomedical Accelerator at Harvard University. Fluorescence microscopy was performed at the Nikon Imaging Center at Harvard Medical School. Electron microscopy was performed at the Harvard Medical School Electron Microscopy Facility.
Publisher Copyright:
© 2018 National Academy of Sciences. All Rights Reserved.
PY - 2018/6/26
Y1 - 2018/6/26
N2 - New drugs are needed to treat gram-negative bacterial infections. These bacteria are protected by an outer membrane which prevents many antibiotics from reaching their cellular targets. The outer leaflet of the outer membrane contains LPS, which is responsible for creating this permeability barrier. Interfering with LPS biogenesis affects bacterial viability. We developed a cell-based screen that identifies inhibitors of LPS biosynthesis and transport by exploiting the nonessentiality of this pathway in Acinetobacter. We used this screen to find an inhibitor of MsbA, an ATP-dependent flippase that translocates LPS across the inner membrane. Treatment with the inhibitor caused mislocalization of LPS to the cell interior. The discovery of an MsbA inhibitor, which is universally conserved in all gram-negative bacteria, validates MsbA as an antibacterial target. Because our cell-based screen reports on the function of the entire LPS biogenesis pathway, it could be used to identify compounds that inhibit other targets in the pathway, which can provide insights into vulnerabilities of the gram-negative cell envelope.
AB - New drugs are needed to treat gram-negative bacterial infections. These bacteria are protected by an outer membrane which prevents many antibiotics from reaching their cellular targets. The outer leaflet of the outer membrane contains LPS, which is responsible for creating this permeability barrier. Interfering with LPS biogenesis affects bacterial viability. We developed a cell-based screen that identifies inhibitors of LPS biosynthesis and transport by exploiting the nonessentiality of this pathway in Acinetobacter. We used this screen to find an inhibitor of MsbA, an ATP-dependent flippase that translocates LPS across the inner membrane. Treatment with the inhibitor caused mislocalization of LPS to the cell interior. The discovery of an MsbA inhibitor, which is universally conserved in all gram-negative bacteria, validates MsbA as an antibacterial target. Because our cell-based screen reports on the function of the entire LPS biogenesis pathway, it could be used to identify compounds that inhibit other targets in the pathway, which can provide insights into vulnerabilities of the gram-negative cell envelope.
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U2 - 10.1073/pnas.1804670115
DO - 10.1073/pnas.1804670115
M3 - Article
C2 - 29735709
AN - SCOPUS:85049052502
SN - 0027-8424
VL - 115
SP - 6834
EP - 6839
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 26
ER -