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Multi-scale model of bacterial cell-cell interactions

Periodic Reporting for period 2 - T6S (Multi-scale model of bacterial cell-cell interactions)

Reporting period: 2018-07-01 to 2019-12-31

Here we address type 6 secretion system function on different scales of resolution. Understanding bacterial secretion systems is relevant for understanding fundamental bacterial cell biology. The objectives are to understand the structure of the macromolecular machine.
Towards Aim 1, we established in situ structures of the T6SS in two different species by sub-tomogram averaging. See Böck et al. Science and Rapisarda et al. EMBO Journal.

Towards Aim 2, we established the mechanism of T6SS bundle formation in Amoebophilus asiaticus (Böck et al., Science). Ongoing efforts address this issue in Pseudomonas aeruginosa. Furthermore, we showed a mechanism for T6SS length regulation in EAEC (Szwedziak and Pilhofer, Nature Communications).

Towards Aim 3, we established a workflow for cryo-focused ion beam milling in order to thin samples for cryo-electron tomography (Medeiros et al., Ultramicroscopy). Subsequently, we showed the suitability of the method to answer biological questions (Cai et al., Molecular Biology of the Cell). We also used the method in order to study the function of the Amoebophilus T6SS (Böck et al., Science). Furthermore, we established cryo-light microscopy for the identification of rare targets (Szwedziak and Pilhofer, Nature Communications).
We envision further methodological advances that will allow us to takle further pressing questions in the field of T6SS-mediated cell-cell interactions.
T6SS membrane complex