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Min protein patterns emerge from rapid rebinding and membrane interaction of MinE

Nature Structural & Molecular Biology volume 18, pages 577–583 (2011)Cite this article

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Abstract

In Escherichia coli, the pole-to-pole oscillation of the Min proteins directs septum formation to midcell, which is required for symmetric cell division. In vitro, protein waves emerge from the self-organization of MinD, a membrane-binding ATPase, and its activator MinE. For wave propagation, the proteins need to cycle through states of collective membrane binding and unbinding. Although MinD presumably undergoes cooperative membrane attachment, it is unclear how synchronous detachment is coordinated. We used confocal and single-molecule microscopy to elucidate the order of events during Min wave propagation. We propose that protein detachment at the rear of the wave, and the formation of the E-ring, are accomplished by two complementary processes: first, local accumulation of MinE due to rapid rebinding, leading to dynamic instability; and second, a structural change induced by membrane-interaction of MinE in an equimolar MinD–MinE (MinDE) complex, which supports the robustness of pattern formation.

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Figure 1: MinD, MinE and eGFP-MinC in traveling waves in vitro.
Figure 2: Dynamics of Min proteins and their relationships.
Figure 3: MinE accumulates at the rear of the wave during wave propagation.
Figure 4: Single-molecule studies on Min proteins.
Figure 5: Membrane binding is not required for MinE accumulation.
Figure 6: Model of Min-protein wave propagation.

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Acknowledgements

We would like to thank D. RayChaudhuri (Tufts University) for plasmid pZH101, J. Howard, S. Vogel and M. Mayer (all Max Planck Institute for Molecular Cell Biology and Genetics) for comments on the manuscript and D. Mullins (University of California, San Francisco) for discussions. M.L. received a scholarship from the Studienstiftung des deutschen Volkes. This work was also supported by the Max Planck Society (M.L., E.F.-F., P.S.).

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Authors and Affiliations

  1. Biophysics, BIOTEC, Dresden University of Technology, Dresden, Germany

    Martin Loose, Christoph Herold & Petra Schwille

  2. Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany

    Martin Loose & Petra Schwille

  3. Max Planck Institute for the Physics of Complex Systems, Dresden, Germany

    Elisabeth Fischer-Friedrich

  4. Theoretische Physik, Universitaet des Saarlandes, Saarbruecken, Germany

    Karsten Kruse

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Contributions

M.L., P.S. and K.K. designed the research, M.L. conducted the research, E.F.-F. and C.H. wrote the tracking software, C.H. built the single-molecule TIRF setup, M.L. analyzed and interpreted the data with the help of all authors, and M.L., E.F.-F. and P.S. wrote the paper with the help of K.K.

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Correspondence to Petra Schwille.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7, Supplementary Methods and Supplementary Discussion (PDF 4480 kb)

Supplementary Video 1

Confocal fluorescence micrographs showing waves of MinD (0.8 μM with 10 mol % MinDCy3), MinE (1.2 μM MinE with 10 mol % MinE-Cy5) and MinC-eGFP (0.02 μM) on a supported lipid membrane. Scale bar is 50 μm. (MOV 3723 kb)

Supplementary Video 2

TIRF micrograph of waves of MinD (0.8 μM, doped with 5 mol % MinD-Alexa 488) and MinE (1.2 μM MinE, doped with 5 mol % MinE-Cy5) middle. The merged channels are shown on the bottom. Scale bar is 10 μm. (MOV 4333 kb)

Supplementary Video 3

Single molecule TIRF micrograph of waves of MinE (1.2 μM, doped with 0.01 mol % MinECy5) (top) and MinD (0.8 μM, doped with 10 mol % MinD-Alexa 488) (middle) and overlay (bottom). Scale bar is 20 μm. (MOV 11536 kb)

Supplementary Video 4

Matlab based particle tracking allowed us to track and analyze single particles inside the waves. The movie shows the calculated trajectories for single MinE proteins. (MOV 5252 kb)

Supplementary Video 5

Confocal fluorescence micrographs showing waves of MinD (blue, 0.8 μM with 10 mol % MinD-Alexa 488), MinE C1 (red, 1.2 μM MinE C1with 10 mol % MinE C1-Cy5). Scale bar is 50 μm. (MOV 2197 kb)

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Loose, M., Fischer-Friedrich, E., Herold, C. et al. Min protein patterns emerge from rapid rebinding and membrane interaction of MinE. Nat Struct Mol Biol 18, 577–583 (2011). https://doi.org/10.1038/nsmb.2037

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