1. Loss of YhcB results in dysregulation of coordinated peptidoglycan, LPS and phospholipid synthesis during Escherichia coli cell growth
- Author
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Emily C. A. Goodall, Georgia L. Isom, Jessica L. Rooke, Karthik Pullela, Christopher Icke, Zihao Yang, Gabriela Boelter, Alun Jones, Isabel Warner, Rochelle Da Costa, Bing Zhang, James Rae, Wee Boon Tan, Matthias Winkle, Antoine Delhaye, Eva Heinz, Jean-Francois Collet, Adam F. Cunningham, Mark A. Blaskovich, Robert G. Parton, Jeff A. Cole, Manuel Banzhaf, Shu-Sin Chng, Waldemar Vollmer, Jack A. Bryant, and Ian R. Henderson
- Subjects
Lipopolysaccharides ,Cancer Research ,GENE-CLUSTER ,Polymers ,LIPOPOLYSACCHARIDE ,QH426-470 ,Biochemistry ,Cell Wall ,Antibiotics ,Mobile Genetic Elements ,Medicine and Health Sciences ,Materials ,Genetics (clinical) ,Phospholipids ,Genetics & Heredity ,0303 health sciences ,Antimicrobials ,Escherichia coli Proteins ,Drugs ,Genomics ,Lipids ,READ ALIGNMENT ,PENICILLIN-BINDING PROTEINS ,Chemistry ,Macromolecules ,Physical Sciences ,qu_450 ,Oxidoreductases ,Life Sciences & Biomedicine ,Cell Division ,Research Article ,Materials Science ,Peptidoglycan ,Library Screening ,Biosynthesis ,Research and Analysis Methods ,Microbiology ,03 medical and health sciences ,Genetic Elements ,Vancomycin ,Microbial Control ,Escherichia coli ,Genetics ,Polymyxins ,Molecular Biology Techniques ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,Pharmacology ,Molecular Biology Assays and Analysis Techniques ,Science & Technology ,COMPLEX ,IDENTIFICATION ,030306 microbiology ,LIPID HOMEOSTASIS ,qw_138 ,Cell Membrane ,Transposable Elements ,Biology and Life Sciences ,Gene Expression Regulation, Bacterial ,Peptidoglycans ,Polymer Chemistry ,TRANSPORT ,Mutagenesis ,OUTER-MEMBRANE ,ENTEROBACTERIAL COMMON ANTIGEN - 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., Author summary All life depends on a cell envelope to enclose the chemical reactions that make life possible. But how do cell envelopes grow? How each component of the cell envelope is incorporated into the envelope at the correct amount, in the correct place, and at the correct time, to prevent cell death, has been a long-standing question in bacteriology. Using a unique combination of high throughput chemical genetic screens we identified yhcB, a conserved gene of unknown function, required for the maintenance of cell envelope integrity in Escherichia coli. Loss of YhcB results in aberrant cell size driven by the production of excess membrane phospholipids. Subsequent molecular and biochemical analyses suggest YhcB influences the spatiotemporal biogenesis of LPS, peptidoglycan and membrane phospholipids. Our data indicate YhcB is a key regulator of cell envelope growth in Gram-negative bacteria playing a crucial role in coordinating cell width, elongation, and division to maintain cell envelope integrity.
- Published
- 2021