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Dynamic Organization of SecA and SecY Secretion Complexes in the B. subtilis Membrane

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  • Alex Dajkovic
  • Elizabeth Hinde
  • Calum MacKichan
  • Rut Carballido-Lopez

Abstract

In prokaryotes, about one third of cellular proteins are translocated across the plasma membrane or inserted into it by concerted action of the cytoplasmic ATPase SecA and the universally conserved SecYEG heterotrimeric polypeptide-translocating pore. Secretion complexes have been reported to localize in specific subcellular sites in Bacillus subtilis. In this work, we used a combination of total internal reflection microscopy, scanning fluorescence correlation spectroscopy, and pair correlation function to study the localization and dynamics of SecA and SecY in growing Bacillus subtilis cells. Both SecA and SecY localized in transient and dynamic foci in the cytoplasmic membrane, which displayed no higher-level organization in helices. Foci of SecA and SecY were in constant flux with freely diffusing SecA and SecY molecules. Scanning FCS confirmed the existence of populations of cellular SecA and SecY molecules with a wide range of diffusion coefficients. Diffusion of SecY as an uncomplexed molecular species was short-lived and only local while SecY complexed with its protein partners traversed distances of over half a micrometer in the cell.

Suggested Citation

  • Alex Dajkovic & Elizabeth Hinde & Calum MacKichan & Rut Carballido-Lopez, 2016. "Dynamic Organization of SecA and SecY Secretion Complexes in the B. subtilis Membrane," PLOS ONE, Public Library of Science, vol. 11(6), pages 1-17, June.
    • Handle: RePEc:plo:pone00:0157899
    DOI: 10.1371/journal.pone.0157899
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    1. Fredric Carlsson & Margaretha Stålhammar-Carlemalm & Klas Flärdh & Charlotta Sandin & Eric Carlemalm & Gunnar Lindahl, 2006. "Signal sequence directs localized secretion of bacterial surface proteins," Nature, Nature, vol. 442(7105), pages 943-946, August.
    2. Tom A. Rapoport, 2007. "Protein translocation across the eukaryotic endoplasmic reticulum and bacterial plasma membranes," Nature, Nature, vol. 450(7170), pages 663-669, November.
    3. Bert van den Berg & William M. Clemons & Ian Collinson & Yorgo Modis & Enno Hartmann & Stephen C. Harrison & Tom A. Rapoport, 2004. "X-ray structure of a protein-conducting channel," Nature, Nature, vol. 427(6969), pages 36-44, January.
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