A5021709138- Bacterial Genetics and Biotechnology
- Escherichia coli research studies
- Bacteriophages and microbial interactions
- Lipid Membrane Structure and Behavior
- Antibiotic Resistance in Bacteria
- RNA and protein synthesis mechanisms
- Mycobacterium research and diagnosis
- Salmonella and Campylobacter epidemiology
- Vibrio bacteria research studies
- Biopolymer Synthesis and Applications
- Metabolism and Genetic Disorders
- Protein Structure and Dynamics
- Tuberculosis Research and Epidemiology
- Neurological diseases and metabolism
- Porphyrin Metabolism and Disorders
- Genomics and Phylogenetic Studies
- Listeria monocytogenes in Food Safety
- Transgenic Plants and Applications
- Immune Response and Inflammation
- RNA Interference and Gene Delivery
- Inflammasome and immune disorders
- Immune responses and vaccinations
- Glycosylation and Glycoproteins Research
- Medical Research and Treatments
- Endoplasmic Reticulum Stress and Disease
University of Cambridge
2017-2024
National Cancer Institute
2023-2024
Center for Cancer Research
2024
Many valuable biopharmaceutical and biotechnological proteins have been produced in Escherichia coli, however these are almost exclusively localised the cytoplasm or periplasm. This presents challenges for purification, i.e. removal of contaminating cellular constituents. One solution is secretion directly into surrounding media, which we achieved via 'hijack' flagellar type III system (FT3SS). Ordinarily subunits exported through centre growing flagellum, before assembly at tip. However,...
Type III Secretion Systems (T3SS) deliver subunits from the bacterial cytosol to nascent cell surface flagella. Early flagellar that form rod and hook substructures are unchaperoned contain their own export signals. A gate recognition motif (GRM) docks them at FlhBc component of FlhAB-FliPQR gate, but must then be opened unfolded pass through channel. This induced us seek further signals on subunits. Here, we identify a second signal extreme N-terminus determine key is its hydrophobicity. We...
Abstract Coenzyme A (CoA) is a fundamental co-factor for all life, involved in numerous metabolic pathways and cellular processes, its biosynthetic pathway has raised substantial interest as drug target against multiple pathogens including Mycobacterium tuberculosis . The biosynthesis of CoA performed five steps, with the second third steps being catalysed vast majority prokaryotes, M. , by single bifunctional protein, CoaBC. Depletion CoaBC was found to be bactericidal Here we report first...
Abstract Bacterial flagellar subunits are exported across the cell membrane by Type III Secretion System (fT3SS), powered proton motive force (pmf) and a specialized ATPase that enables export gate to utilize pmf electric potential (ΔΨ). Export activation is mediated stalk, FliJ, but how this process regulated prevent wasteful dissipation of in absence subunit cargo not known. Here, we show FliJ chaperones. binds unladen chaperones and, using novel chaperone variants specifically defective...
Abstract Translational control in pathogenic bacteria is fundamental to gene expression and affects virulence other infection phenotypes. We used an enhanced ribosome profiling protocol coupled with parallel transcriptomics capture accurately the global translatome of two evolutionarily distant bacteria—the Gram-negative bacterium Salmonella Gram-positive Listeria . find that use different mechanisms translationally regulate protein synthesis. In , addition expected correlation between...
Abstract During bacterial infection both the host cell and its invader must divert intracellular resources to synthesise specific proteins in a timely manner. For host, these factors may be needed for innate immune responses, including programmed death, bacteria newly synthesized survival counteract responses. Salmonella is an important food-borne pathogen that invades multiplies within cells. It well established invasion of epithelial cells dependent upon SPI-1 Type III injectisome,...
Type III Secretion Systems (T3SS) transport proteins from the bacterial cytosol for assembly into cell surface nanomachines or direct delivery target eukaryotic cells. At core of flagellar T3SS, FlhAB-FliPQR export gate regulates protein entry channel whilst maintaining integrity membrane. Here, we identify critical residues in FliR plug that stabilise closed conformation, preserving membrane permeability barrier, and show opens closes response to substrate availability. Our data indicate...
Abstract Translational control in pathogenic bacteria is fundamental to gene expression and affects virulence other infection phenotypes. We used an enhanced ribosome profiling protocol coupled with parallel transcriptomics capture accurately the global translatome of two evolutionarily distant – Gram-negative bacterium Salmonella Gram positive Listeria find that use different mechanisms translationally regulate protein synthesis. In Salmonella, addition expected correlation between...
ABSTRACT Type III secretion systems (T3SSs) are essential for motility and virulence in many bacterial pathogens. Proteins destined the flagellar T3SS contain at least two export signals their N-terminal D 0 domain. Here, we describe a third carboxy (C)-terminal signal early subunits that facilitates subunit targeting to machinery. Mutational analysis identified critical residues within hook C-terminal signal. The ATPase cytoplasmic ring components were not required this targeting,...
Abstract Coenzyme A (CoA) is a fundamental co-factor for all life, involved in numerous metabolic pathways and cellular processes, its biosynthetic pathway has raised substantial interest as drug target against multiple pathogens including Mycobacterium tuberculosis . The biosynthesis of CoA performed five steps, with the second third steps being catalysed vast majority prokaryotes, M. , by single bifunctional protein, CoaBC. Depletion CoaBC was found to be bactericidal Here we report first...
Abstract Type III Secretion Systems (T3SS) deliver subunits from the bacterial cytosol to nascent cell surface flagella. Early flagellar that form rod and hook substructures are unchaperoned contain their own export signals. A gate recognition motif (GRM) docks them at FlhBc component of FlhAB-FliPQR gate, but must then be opened unfolded pass through channel. This induced us seek further signals on subunits. Here, we identify a second signal extreme N-terminus determine key is its...
Abstract Bacterial flagellar subunits are exported across the cell membrane by Type III Secretion System (fT3SS), powered proton motive force (pmf) and a specialized ATPase that enables export gate to utilise pmf electric potential (ΔΨ). Export activation is mediated stalk, FliJ, but how this process regulated prevent wasteful dissipation of in absence subunit cargo not known. Here, we show FliJ chaperones. binds unladen chaperones and, using novel chaperone variants specifically defective...
Bacterial flagella are assembled from thousands of protein subunits that unfolded and exported via a specialized type III secretion system. Subunit export is fuelled by the proton motive force (PMF) facilitated cytoplasmic ATPase complex comprising FliH, FliI FliJ, which evolutionarily related to components F1 ATPase. The FliJ stalk component binds gate FlhA, allowing it utilise ΔΨ drive highly efficient subunit export. What unclear how activation FlhA regulated prevent constitutive influx...
Abstract Type III Secretion Systems (T3SS) transport proteins from the bacterial cytosol for assembly into cell surface nanomachines or direct delivery target eukaryotic cells. At core of flagellar T3SS, FlhAB-FliPQR export gate regulates protein entry channel whilst maintaining integrity membrane. Here, we identify critical residues in FliR plug that stabilise closed conformation, preserving membrane permeability barrier, and show opens closes response to substrate availability. Our data...