Abstract
The cell envelope is essential for viability in all domains of life. It retains enzymes and substrates within a confined space while providing a protective barrier to the external environment. Destabilising the envelope of bacterial pathogens is a common strategy employed by antimicrobial treatment. However, even in one of the best studied organisms, Escherichia coli, there remain gaps in our understanding of how the synthesis of the successive layers of the cell envelope are coordinated during growth and cell division. Here, we used a whole-genome phenotypic screen to identify mutants with a defective cell envelope. We report that loss of yhcB, a conserved gene of unknown function, results in loss of envelope stability, increased cell permeability and dysregulated control of cell size. Using whole genome transposon mutagenesis strategies, we report the comprehensive genetic interaction network of yhcB, revealing all genes with a synthetic negative and a synthetic positive relationship. These genes include those previously reported to have a role in cell envelope biogenesis. Surprisingly, we identified genes previously annotated as essential that became non-essential in a ΔyhcB background. Subsequent analyses suggest that YhcB functions at the junction of several envelope biosynthetic pathways coordinating the spatiotemporal growth of the cell, highlighting YhcB as an as yet unexplored antimicrobial target.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cell Division / genetics
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Cell Membrane / genetics
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Cell Membrane / microbiology
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Cell Wall / genetics*
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Cell Wall / microbiology
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Escherichia coli / genetics
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Escherichia coli Proteins / genetics*
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Gene Expression Regulation, Bacterial / genetics
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Lipopolysaccharides / biosynthesis
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Lipopolysaccharides / genetics*
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Mutagenesis
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Oxidoreductases / genetics*
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Peptidoglycan / genetics*
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Phospholipids / biosynthesis
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Phospholipids / genetics
Substances
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Escherichia coli Proteins
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Lipopolysaccharides
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Peptidoglycan
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Phospholipids
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Oxidoreductases
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YhcB protein, E coli
Grants and funding
ECAG was supported by a Biotechnology and Biological Sciences Research Council scholarship to ECAG as part of the Midlands Integrative Biosciences Training Partnership IRH. The research was supported by a European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 721484 awarded to WV and IRH. B.Z. was supported by a CSC-UQ PhD Scholarship (China Scholarship Council) to MB. This work was also supported by the National Health and Medical Research Council of Australia (grants APP1140064 and APP1150083 and fellowship APP1156489 to R.G.P.). RGP is supported by the Australian Research Council (ARC) Centre of Excellence in Convergent Bio-Nano Science and Technology CE140100036. This work was supported by the University of Queensland funding to IRH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.