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Title: The Lipid A 1-Phosphatase, LpxE, Functionally Connects Multiple Layers of Bacterial Envelope Biogenesis

Abstract

Although distinct lipid phosphatases are thought to be required for processing lipid A (component of the outer leaflet of the outer membrane), glycerophospholipid (component of the inner membrane and the inner leaflet of the outer membrane), and undecaprenyl pyrophosphate (C 55-PP; precursors of peptidoglycan and O antigens of lipopolysaccharide) in Gram-negative bacteria, we report that the lipid A 1-phosphatases, LpxEs, functionally connect multiple layers of cell envelope biogenesis in Gram-negative bacteria. We found that Aquifex aeolicus LpxE structurally resembles YodM in Bacillus subtilis, a phosphatase for phosphatidylglycerol phosphate (PGP) with a weak in vitro activity on C 55-PP, and rescues Escherichia coli deficient in PGP and C 55-PP phosphatase activities; deletion of lpxE in Francisella novicida reduces the MIC value of bacitracin, indicating a significant contribution of LpxE to the native bacterial C 55-PP phosphatase activity. Suppression of plasmid-borne lpxE in F. novicida deficient in chromosomally encoded C 55-PP phosphatase activities results in cell enlargement, loss of O-antigen repeats of lipopolysaccharide, and ultimately cell death. These discoveries implicate LpxE as the first example of a multifunctional regulatory enzyme that orchestrates lipid A modification, O-antigen production, and peptidoglycan biogenesis to remodel multiple layers of the Gram-negative bacterial envelope.

Authors:
 [1];  [2];  [2];  [1];  [1];  [1];  [3];  [1]; ORCiD logo [1]; ORCiD logo [2]; ;
  1. Duke Univ. Medical Center, Durham, NC (United States)
  2. Korea Inst. of Science and Technology (KIST), Seoul (Republic of Korea); Korea Univ. of Science and Technology, Seoul (Republic of Korea)
  3. Korea Inst. of Science and Technology (KIST), Seoul (Republic of Korea)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
Intramural Research Program of Korea Institute of Science and Technology (KIST); National Research Foundation of Korea (NRF); National Institutes of Health (NIH); National Institute of General Medical Sciences (NIGMS); USDOE Office of Science (SC)
OSTI Identifier:
1545868
Grant/Contract Number:  
[2018R1A2B2008995; GM51310; GM115355; P30 GM124165; S10 RR029205; AC02-06CH11357]
Resource Type:
Accepted Manuscript
Journal Name:
mBio (Online)
Additional Journal Information:
[Journal Name: mBio (Online); Journal Volume: 10; Journal Issue: 3]; Journal ID: ISSN 2150-7511
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; bacterial cell envelope biogenesis; lipid A 1-phosphate phosphatase; phosphatidylglycerol phosphate phosphatase; type 2 phosphatidic acid phosphatase (PAP2) superfamily; undecaprenyl pyrophosphate phosphatase

Citation Formats

Zhao, Jinshi, An, Jinsu, Hwang, Dohyeon, Wu, Qinglin, Wang, Su, Gillespie, Robert A., Yang, Eun Gyeong, Guan, Ziqiang, Zhou, Pei, Chung, Hak Suk, Stephen Trent, M., and Salama, Nina R. The Lipid A 1-Phosphatase, LpxE, Functionally Connects Multiple Layers of Bacterial Envelope Biogenesis. United States: N. p., 2019. Web. doi:10.1128/mBio.00886-19.
Zhao, Jinshi, An, Jinsu, Hwang, Dohyeon, Wu, Qinglin, Wang, Su, Gillespie, Robert A., Yang, Eun Gyeong, Guan, Ziqiang, Zhou, Pei, Chung, Hak Suk, Stephen Trent, M., & Salama, Nina R. The Lipid A 1-Phosphatase, LpxE, Functionally Connects Multiple Layers of Bacterial Envelope Biogenesis. United States. doi:10.1128/mBio.00886-19.
Zhao, Jinshi, An, Jinsu, Hwang, Dohyeon, Wu, Qinglin, Wang, Su, Gillespie, Robert A., Yang, Eun Gyeong, Guan, Ziqiang, Zhou, Pei, Chung, Hak Suk, Stephen Trent, M., and Salama, Nina R. Tue . "The Lipid A 1-Phosphatase, LpxE, Functionally Connects Multiple Layers of Bacterial Envelope Biogenesis". United States. doi:10.1128/mBio.00886-19. https://www.osti.gov/servlets/purl/1545868.
@article{osti_1545868,
title = {The Lipid A 1-Phosphatase, LpxE, Functionally Connects Multiple Layers of Bacterial Envelope Biogenesis},
author = {Zhao, Jinshi and An, Jinsu and Hwang, Dohyeon and Wu, Qinglin and Wang, Su and Gillespie, Robert A. and Yang, Eun Gyeong and Guan, Ziqiang and Zhou, Pei and Chung, Hak Suk and Stephen Trent, M. and Salama, Nina R.},
abstractNote = {Although distinct lipid phosphatases are thought to be required for processing lipid A (component of the outer leaflet of the outer membrane), glycerophospholipid (component of the inner membrane and the inner leaflet of the outer membrane), and undecaprenyl pyrophosphate (C55-PP; precursors of peptidoglycan and O antigens of lipopolysaccharide) in Gram-negative bacteria, we report that the lipid A 1-phosphatases, LpxEs, functionally connect multiple layers of cell envelope biogenesis in Gram-negative bacteria. We found that Aquifex aeolicus LpxE structurally resembles YodM in Bacillus subtilis, a phosphatase for phosphatidylglycerol phosphate (PGP) with a weak in vitro activity on C55-PP, and rescues Escherichia coli deficient in PGP and C55-PP phosphatase activities; deletion of lpxE in Francisella novicida reduces the MIC value of bacitracin, indicating a significant contribution of LpxE to the native bacterial C55-PP phosphatase activity. Suppression of plasmid-borne lpxE in F. novicida deficient in chromosomally encoded C55-PP phosphatase activities results in cell enlargement, loss of O-antigen repeats of lipopolysaccharide, and ultimately cell death. These discoveries implicate LpxE as the first example of a multifunctional regulatory enzyme that orchestrates lipid A modification, O-antigen production, and peptidoglycan biogenesis to remodel multiple layers of the Gram-negative bacterial envelope.},
doi = {10.1128/mBio.00886-19},
journal = {mBio (Online)},
number = [3],
volume = [10],
place = {United States},
year = {2019},
month = {6}
}

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Works referenced in this record:

Crystal structure of lipid phosphatase Escherichia coli phosphatidylglycerophosphate phosphatase B
journal, May 2014

  • Fan, J.; Jiang, D.; Zhao, Y.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 21
  • DOI: 10.1073/pnas.1403097111

Identification of Multiple Genes Encoding Membrane Proteins with Undecaprenyl Pyrophosphate Phosphatase (UppP) Activity in Escherichia coli
journal, March 2005

  • Ghachi, Meriem El; Derbise, Anne; Bouhss, Ahmed
  • Journal of Biological Chemistry, Vol. 280, Issue 19
  • DOI: 10.1074/jbc.M412277200

Curative Treatment of Severe Gram-Negative Bacterial Infections by a New Class of Antibiotics Targeting LpxC
journal, July 2017


Secretory Production of Recombinant Proteins in Escherichia coli
journal, January 2010


The Lipid A 1-Phosphatase of Helicobacter pylori Is Required for Resistance to the Antimicrobial Peptide Polymyxin
journal, June 2006

  • Tran, A. X.; Whittimore, J. D.; Wyrick, P. B.
  • Journal of Bacteriology, Vol. 188, Issue 12
  • DOI: 10.1128/JB.00146-06

Helicobacter pylori versus the Host: Remodeling of the Bacterial Outer Membrane Is Required for Survival in the Gastric Mucosa
journal, December 2011


Structural Insight into Substrate Selection and Catalysis of Lipid Phosphate Phosphatase PgpB in the Cell Membrane
journal, July 2016

  • Tong, Shuilong; Lin, Yibin; Lu, Shuo
  • Journal of Biological Chemistry, Vol. 291, Issue 35
  • DOI: 10.1074/jbc.M116.737874

Structure, inhibition, and regulation of essential lipid A enzymes
journal, November 2017

  • Zhou, Pei; Zhao, Jinshi
  • Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1862, Issue 11
  • DOI: 10.1016/j.bbalip.2016.11.014

MsbA Transporter-dependent Lipid A 1-Dephosphorylation on the Periplasmic Surface of the Inner Membrane: TOPOGRAPHY OF
journal, August 2004

  • Wang, Xiaoyuan; Karbarz, Mark J.; McGrath, Sara C.
  • Journal of Biological Chemistry, Vol. 279, Issue 47
  • DOI: 10.1074/jbc.M409078200

Altered lipid A structures and polymyxin hypersensitivity of Rhizobium etli mutants lacking the LpxE and LpxF phosphatases
journal, May 2010

  • Ingram, Brian O.; Sohlenkamp, Christian; Geiger, Otto
  • Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1801, Issue 5
  • DOI: 10.1016/j.bbalip.2010.02.001

Purification and Characterization of the Lipid A 1-Phosphatase LpxE of Rhizobium leguminosarum
journal, November 2008

  • Karbarz, Mark J.; Six, David A.; Raetz, Christian R. H.
  • Journal of Biological Chemistry, Vol. 284, Issue 1
  • DOI: 10.1074/jbc.M808390200

Phosphatidate phosphatases and diacylglycerol pyrophosphate phosphatases in Saccharomyces cerevisiae and Escherichia coli
journal, September 1997


Crystal structure and biochemical characterization of the transmembrane PAP2 type phosphatidylglycerol phosphate phosphatase from Bacillus subtilis
journal, February 2017

  • Ghachi, Meriem El; Howe, Nicole; Auger, Rodolphe
  • Cellular and Molecular Life Sciences, Vol. 74, Issue 12
  • DOI: 10.1007/s00018-017-2464-6

Structure and Biosynthesis of Free Lipid A Molecules That Replace Lipopolysaccharide in Francisella tularensis subsp. novicida
journal, December 2006

  • Wang, Xiaoyuan; Ribeiro, Anthony A.; Guan, Ziqiang
  • Biochemistry, Vol. 45, Issue 48
  • DOI: 10.1021/bi061767s

The structural basis of lipopolysaccharide recognition by the TLR4–MD-2 complex
journal, March 2009

  • Park, Beom Seok; Song, Dong Hyun; Kim, Ho Min
  • Nature, Vol. 458, Issue 7242
  • DOI: 10.1038/nature07830

Drug design from the cryptic inhibitor envelope
journal, February 2016

  • Lee, Chul-Jin; Liang, Xiaofei; Wu, Qinglin
  • Nature Communications, Vol. 7, Issue 1
  • DOI: 10.1038/ncomms10638

Lipopolysaccharide Endotoxins
journal, June 2002


Gapped BLAST and PSI-BLAST: a new generation of protein database search programs
journal, September 1997

  • Altschul, Stephen F.; Madden, Thomas L.; Schäffer, Alejandro A.
  • Nucleic Acids Research, Vol. 25, Issue 17, p. 3389-3402
  • DOI: 10.1093/nar/25.17.3389

The Escherichia coli pgpB Gene Encodes for a Diacylglycerol Pyrophosphate Phosphatase Activity
journal, November 1996

  • Dillon, Deirdre A.; Wu, Wen-I; Riedel, Bettina
  • Journal of Biological Chemistry, Vol. 271, Issue 48
  • DOI: 10.1074/jbc.271.48.30548

LpxI structures reveal how a lipid A precursor is synthesized
journal, October 2012

  • Metzger, Louis E.; Lee, John K.; Finer-Moore, Janet S.
  • Nature Structural & Molecular Biology, Vol. 19, Issue 11
  • DOI: 10.1038/nsmb.2393

Three Phosphatidylglycerol-phosphate Phosphatases in the Inner Membrane of Escherichia coli
journal, December 2010

  • Lu, Yi-Hsueh; Guan, Ziqiang; Zhao, Jinshi
  • Journal of Biological Chemistry, Vol. 286, Issue 7
  • DOI: 10.1074/jbc.M110.199265

Lipid A Modification Systems in Gram-Negative Bacteria
journal, June 2007


Identification of a novel phosphatase sequence motif
journal, February 1997


Biosynthesis of Undecaprenyl Phosphate-Galactosamine and Undecaprenyl Phosphate-Glucose in Francisella novicida
journal, February 2009

  • Song, Feng; Guan, Ziqiang; Raetz, Christian R. H.
  • Biochemistry, Vol. 48, Issue 6
  • DOI: 10.1021/bi802212t

One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products
journal, May 2000

  • Datsenko, K. A.; Wanner, B. L.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 12, p. 6640-6645
  • DOI: 10.1073/pnas.120163297

Constitutive versus thermoinducible expression of heterologous proteins inEscherichia coli based on strong PR,PL promoters from phage lambda
journal, May 2003

  • Menart, Viktor; Jev?evar, Simona; Vilar, Mateja
  • Biotechnology and Bioengineering, Vol. 83, Issue 2
  • DOI: 10.1002/bit.10660

Inflammatory caspases are innate immune receptors for intracellular LPS
journal, August 2014


A comprehensive transposon mutant library of Francisella novicida , a bioweapon surrogate
journal, January 2007

  • Gallagher, Larry A.; Ramage, Elizabeth; Jacobs, Michael A.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 3
  • DOI: 10.1073/pnas.0606713104

Integral membrane lipid phosphatases/phosphotransferases: common structure and diverse functions
journal, April 2005

  • Sigal, Yury J.; McDERMOTT, Mark I.; Morris, Andrew J.
  • Biochemical Journal, Vol. 387, Issue 2
  • DOI: 10.1042/BJ20041771

A Francisella Mutant in Lipid A Carbohydrate Modification Elicits Protective Immunity
journal, January 2008


Mutations of Francisella novicida that Alter the Mechanism of Its Phagocytosis by Murine Macrophages
journal, July 2010


An Alternative Route for UDP-Diacylglucosamine Hydrolysis in Bacterial Lipid A Biosynthesis
journal, August 2010

  • Metzger, Louis E.; Raetz, Christian R. H.
  • Biochemistry, Vol. 49, Issue 31
  • DOI: 10.1021/bi1008744