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Abstract
Here, we describe two new heat shock proteins involved in the assembly of LPS in Escherichia coli, LapA and LapB (lipopolysaccharide assembly protein A and B). lapB mutants were identified based on an increased envelope stress response. Envelope stress responsive pathways control key steps in LPS biogenesis and respond to defects in the LPS assembly. Accordingly, the LPS content in ΔlapB or Δ(lapA lapB) mutants was elevated, with an enrichment of LPS derivatives with truncations in the core region, some of which were pentaacylated and exhibited carbon chain polymorphism. Further, the levels of LpxC, the enzyme that catalyzes the first committed step of lipid A synthesis, were highly elevated in the Δ(lapA lapB) mutant. Δ(lapA lapB) mutant accumulated extragenic suppressors that mapped either to lpxC, waaC, gmhA, or the waaQ operon (LPS biosynthesis) and lpp (Braun′s lipoprotein). Increased synthesis of either FabZ (3-R-hydroxymyristoyl acyl carrier protein dehydratase), slrA (novel RpoE-regulated non-coding sRNA), lipoprotein YceK, toxin HicA, or MurA (UDP-N-acetylglucosamine 1-carboxyvinyltransferase) suppressed some of the Δ(lapA lapB) defects. LapB contains six tetratricopeptide repeats and at the C-terminal end a rubredoxin-like domain that was found to be essential for its activity. In pull-down experiments, LapA and LapB co-purified with LPS, Lpt proteins, FtsH (protease), DnaK and DnaJ (chaperones). A specific interaction was also observed between WaaC and LapB. Our data suggest that LapB coordinates assembly of proteins involved in LPS synthesis at the plasma membrane and regulates turnover of LpxC thereby ensuring balanced biosynthesis of LPS and phospholipids consistent with its essentiality.
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Authors (5)
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Buko Lindner
- Research Center Borstel Leibniz-Center for Medicine and Biosciences
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- Accepted or Published Version
- DOI:
- Digital Object Identifier (open in new tab) 10.1074/jbc.M113.539494
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- Category:
- Articles
- Type:
- artykuł w czasopiśmie wyróżnionym w JCR
- Published in:
-
JOURNAL OF BIOLOGICAL CHEMISTRY
no. 289,
pages 14829 - 14853,
ISSN: 0021-9258 - Language:
- English
- Publication year:
- 2014
- Bibliographic description:
- Klein-Raina G., Kobylak N., Lindner B., Stupak A., Raina S.: Assembly of Lipopolysaccharide in Escherichia coli Requires the Essential LapB Heat Shock Protein// JOURNAL OF BIOLOGICAL CHEMISTRY. -Vol. 289, iss. 21 (2014), s.14829-14853
- DOI:
- Digital Object Identifier (open in new tab) 10.1074/jbc.m113.539494
- Verified by:
- Gdańsk University of Technology
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