Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum
SpoIIIE is a membrane-anchored DNA translocase that localizes to the septal midpoint to mediate chromosome translocation and membrane fission during Bacillus subtilis sporulation. Here we use cell-specific protein degradation and quantitative photoactivated localization microscopy in strains with a thick sporulation septum to investigate the architecture and function of the SpoIIIE DNA translocation complex in vivo. We were able to visualize SpoIIIE complexes with approximately equal numbers of molecules in the mother cell and the forespore. Cell-specific protein degradation showed that only the mother cell complex is required to translocate DNA into the forespore, whereas degradation in either cell reverses membrane fission. Our data suggest that SpoIIIE assembles a coaxially paired channel for each chromosome arm comprised of one hexamer in each cell to maintain membrane fission during DNA translocation. We show that SpoIIIE can operate, in principle, as a bi-directional motor that exports DNA.
- Howard Hughes Medical Inst., Chevy Chase, MD (United States); Univ. of California, Berkeley, CA (United States)
- Univ. of California, San Diego, CA (United States). Div. of Biological Sciences
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- Published Article
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- Additional Journal Information:
- Journal Volume: 4; Journal ID: ISSN 2050-084X
- eLife Sciences Publications, Ltd.
- Research Org:
- Univ. of California, Berkeley, CA (United States)
- Sponsoring Org:
- USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Materials Sciences and Engineering Division; National Institutes of Health (NIH); USDOE
- Country of Publication:
- United States
- 59 BASIC BIOLOGICAL SCIENCES
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