Morphological plasticity of bacteria to stressful conditions
May 2017 – May 2020
Group and collaboration
Ariane Briegel & Dennis Claessen
PhD student: Eveline Ultee
Bacteria use their rigid outer layer to withstand a large variety of external stresses. This outer layer is termed the cell wall: a stress-bearing structure, composed of a cross-linked network of peptidoglycan. Peptidoglycan has proven to be a successful target for antibacterial compounds and for recognition of bacterial pathogens by the host immune system. A proposed mechanism to evade these conditions and survive is the modification or even loss of the cell wall. Research has shown that bacteria can chemically convert into a wall-less or L-form state, in which the cells are able to grow and propagate without a cell wall. This process has been implied in the establishment of persistent infections in pathogenic species such as Mycobacterium tuberculosis. To gain detailed insight into the detailed characteristics of such cell-wall less states, we utilize the related, non-pathogenic Actinomycetales.
Our lab has shown that a selection of filamentous Actinomycetes species is able to spontaneously extrude wall-less bacterial cells upon exposure to hyper osmotic stress (S-cells). In order to understand the structural changes that accompany the formation of wall-less bacteria, we first need to understand the architecture of the cell wall structure of filamentous Actinomycetes itself. In filamentous Actinomycetes, the cell wall synthesis machinery is guided to the tip of the hyphal cells by a multi-protein complex. From the hyphal tip, new peptidoglycan strands are incorporated into the cell wall, which allows the cell to grow and elongate at the pole. Interestingly, the wall-less bacteria extrude from this hyphal tip when exposed to osmotic stress. Studying the cell wall structure and formation and extrusion of wall-less bacteria will contribute to a better understanding of how bacteria can adapt to changing environments and evade antibacterial compounds.
Aims & tasks
- To study the structure of the cell wall in polar growing bacteria
- We will use cryo-electron tomography the visualize isolated sacculi (cell wall material) of model Actinomycete Streptomyces coelicolor and compare new (cell pole) and older regions (lateral sides) of the cell wall
- To understand how the bacterial cell and its cell wall restructure in order to extrude wall-less bacteria.
- We will use a combination of imaging techniques (time-lapse microscopy, (cryo-) fluorescence light and electron microscopy/tomography) to describe structural changes during the formation of wall-less bacteria
- We will analyze the role of members of the multi-protein complex at the tip, which interacts with the peptidoglycan-synthesis machinery