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β-Barrel proteins tether the outer membrane in many Gram-negative bacteria

Journal Article · · Nature Microbiology
 [1];  [1];  [1];  [2];  [2];  [3];  [4];  [5];  [1];  [5];  [4];  [2];  [1]
  1. National Inst. of Health (NIH), Hamilton, MT (United States). National Inst. of Allergy and Infectious Diseases (NIAID)
  2. Univ. of Sheffield (United Kingdom)
  3. Protein Metrics Inc., Cupertino, CA (United States)
  4. Georgia Inst. of Technology, Atlanta, GA (United States)
  5. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)
+ Show Author Affiliations
Gram-negative bacteria have a cell envelope that comprises an outer membrane (OM), a peptidoglycan (PG) layer and an inner membrane (IM). The OM and PG are load-bearing, selectively permeable structures that are stabilized by cooperative interactions between IM and OM proteins. In Escherichia coli, Braun’s lipoprotein (Lpp) forms the only covalent tether between the OM and PG and is crucial for cell envelope stability; however, most other Gram-negative bacteria lack Lpp so it has been assumed that alternative mechanisms of OM stabilization are present. In this work we used a glycoproteomic analysis of PG to show that β-barrel OM proteins are covalently attached to PG in several Gram-negative species, including Coxiella burnetii, Agrobacterium tumefaciens and Legionella pneumophila. In C. burnetii, we found that four different types of covalent attachments occur between OM proteins and PG, with tethering of the β-barrel OM protein BbpA becoming most abundant in the stationary phase and tethering of the lipoprotein LimB similar throughout the cell cycle. Using a genetic approach, we demonstrate that the cell cycle-dependent tethering of BbpA is partly dependent on a developmentally regulated L,D-transpeptidase (Ldt). We use our findings to propose a model of Gram-negative cell envelope stabilization that includes cell cycle control and an expanded role for Ldts in covalently attaching surface proteins to PG.
Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). Compute and Data Environment for Science (CADES)
Sponsoring Organization:
Biotechnology and Biological Sciences Research Council (BBSRC); Medical Research Council (MRC); National Institute of Allergy and Infectious Diseases (NIAID); National Institutes of Health (NIH); National Science Foundation (NSF); USDOE; USDOE Laboratory Directed Research and Development (LDRD) Program
Grant/Contract Number:
AC05-00OR22725
OSTI ID:
1706272
Journal Information:
Nature Microbiology, Journal Name: Nature Microbiology Journal Issue: 1 Vol. 6; ISSN 2058-5276
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English

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59 BASIC BIOLOGICAL SCIENCES
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