A View to a Kill: T6SS-Mediated Cell Killing Visualized by Fluorescence Microscopy

ORAL

Abstract

The Type Six Secretion System (T6SS) is a bacterial toxin-delivery system targeting bacterial cells which neighbor the donor, promoting recipient cell death. The T6SS is widely conserved among Gram-negative bacteria and may be a central determinant in bacterial fitness in polymicrobial communities of particular relevance to chronic infection. Sequence homology of secretion system components to the T4 bacteriophage tail spike, cryoEM reconstructions of the secretion system and fluorescence imaging are all consistent with a dynamic mechanism of secretion. The complex system, which is composed of at least 15 proteins, forms a puncturing apparatus/delivery system which uses a donor protein filament to puncture the recipient cell wall to deliver protein toxins. Using quantitative imaging analysis of multiple fluorescent fusions, we present a detailed characterization of T6SS system dynamics visualized in single cells in multiple bacterial species, developing a model of T6SS function. We present quantitative measurements of the dynamics of the secretion system - from the assembly to contraction to disassembly - in conjunction with quantitative measures of system function, including recipient cell lysis.

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Authors

  • Jacqueline Corbitt

    University of Washington

  • William H. Dowd

    Oregon State University, University of Washington, University of Hong Kong, University of Tennessee, Oak Ridge National Laboratory, University of British Columbia, Univ of Washington, Univ of Cambridge, Michigan State University, Universit\'e de Caen, Argonne National Laboratory, Texas A\&M University -Commerce, Texas A\&M University, Department of Physics, University of Idaho, Moscow, Idaho 83844, USA, Universidad de Salamanca, E-37008 Salamanca, Spain, Department of Physics, Florida State University, Tallahassee, Florida 32306, USA, University of Cambridge, Institute of Astronomy, University of Cambridge, Canadian Institute for Theoretical Astrophysics, University of Toronto, Seoul National University, Chungnam National University, Department of Physics, Oregon State University, TRIUMF, Roosevelt High School, Department of Physics, University of Washington, Oregon State Department of Chemistry, Oregon State School of Electrical Engineering and Computer Science, Oregon State Department of Physics, University of Idaho, Lawrence Livermore National Laboratory, Idaho National Laboratory, Pacific Northwest National Laboratory, Carnegie Institution of Washington, Department of Chemistry, Oregon State University, School of Electrical Engineering and Computer Science, Oregon State University, Washington State University, Harvard University, Idaho Accelerator Center, Idaho Accelerator Center, Idaho State University, 1500 Alvin Ricken drive, Pocatello, ID 83201, USA, CENPA, University of Washington, Physics Division, ANL, NSCL, Michigan State University, Division, ANL, LPC, CAEN, France, Paul Scherrer Institute, US Geological Survey, 12201 Sunrise Valley Drive, Reston, VA, 20192, USA, University of Calgary Department of Physics and Astronomy, University of Calgary Department of Geoscience, None, University of the Fraser Valley, Univ of California, Berkeley, Simon Fraser University, Los Alamos Natl. Lab., University of Science and Technology of China, Hefei, U of Washington, Georgia Institute of Technology, Washington State Univ, National Institute of Standards and Technology and University of Maryland, American University

  • William H. Dowd

    Oregon State University, University of Washington, University of Hong Kong, University of Tennessee, Oak Ridge National Laboratory, University of British Columbia, Univ of Washington, Univ of Cambridge, Michigan State University, Universit\'e de Caen, Argonne National Laboratory, Texas A\&M University -Commerce, Texas A\&M University, Department of Physics, University of Idaho, Moscow, Idaho 83844, USA, Universidad de Salamanca, E-37008 Salamanca, Spain, Department of Physics, Florida State University, Tallahassee, Florida 32306, USA, University of Cambridge, Institute of Astronomy, University of Cambridge, Canadian Institute for Theoretical Astrophysics, University of Toronto, Seoul National University, Chungnam National University, Department of Physics, Oregon State University, TRIUMF, Roosevelt High School, Department of Physics, University of Washington, Oregon State Department of Chemistry, Oregon State School of Electrical Engineering and Computer Science, Oregon State Department of Physics, University of Idaho, Lawrence Livermore National Laboratory, Idaho National Laboratory, Pacific Northwest National Laboratory, Carnegie Institution of Washington, Department of Chemistry, Oregon State University, School of Electrical Engineering and Computer Science, Oregon State University, Washington State University, Harvard University, Idaho Accelerator Center, Idaho Accelerator Center, Idaho State University, 1500 Alvin Ricken drive, Pocatello, ID 83201, USA, CENPA, University of Washington, Physics Division, ANL, NSCL, Michigan State University, Division, ANL, LPC, CAEN, France, Paul Scherrer Institute, US Geological Survey, 12201 Sunrise Valley Drive, Reston, VA, 20192, USA, University of Calgary Department of Physics and Astronomy, University of Calgary Department of Geoscience, None, University of the Fraser Valley, Univ of California, Berkeley, Simon Fraser University, Los Alamos Natl. Lab., University of Science and Technology of China, Hefei, U of Washington, Georgia Institute of Technology, Washington State Univ, National Institute of Standards and Technology and University of Maryland, American University

  • William H. Dowd

    Oregon State University, University of Washington, University of Hong Kong, University of Tennessee, Oak Ridge National Laboratory, University of British Columbia, Univ of Washington, Univ of Cambridge, Michigan State University, Universit\'e de Caen, Argonne National Laboratory, Texas A\&M University -Commerce, Texas A\&M University, Department of Physics, University of Idaho, Moscow, Idaho 83844, USA, Universidad de Salamanca, E-37008 Salamanca, Spain, Department of Physics, Florida State University, Tallahassee, Florida 32306, USA, University of Cambridge, Institute of Astronomy, University of Cambridge, Canadian Institute for Theoretical Astrophysics, University of Toronto, Seoul National University, Chungnam National University, Department of Physics, Oregon State University, TRIUMF, Roosevelt High School, Department of Physics, University of Washington, Oregon State Department of Chemistry, Oregon State School of Electrical Engineering and Computer Science, Oregon State Department of Physics, University of Idaho, Lawrence Livermore National Laboratory, Idaho National Laboratory, Pacific Northwest National Laboratory, Carnegie Institution of Washington, Department of Chemistry, Oregon State University, School of Electrical Engineering and Computer Science, Oregon State University, Washington State University, Harvard University, Idaho Accelerator Center, Idaho Accelerator Center, Idaho State University, 1500 Alvin Ricken drive, Pocatello, ID 83201, USA, CENPA, University of Washington, Physics Division, ANL, NSCL, Michigan State University, Division, ANL, LPC, CAEN, France, Paul Scherrer Institute, US Geological Survey, 12201 Sunrise Valley Drive, Reston, VA, 20192, USA, University of Calgary Department of Physics and Astronomy, University of Calgary Department of Geoscience, None, University of the Fraser Valley, Univ of California, Berkeley, Simon Fraser University, Los Alamos Natl. Lab., University of Science and Technology of China, Hefei, U of Washington, Georgia Institute of Technology, Washington State Univ, National Institute of Standards and Technology and University of Maryland, American University