Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum
Abstract
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.
- Authors:
-
- Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States
- Division of Biological Sciences, University of California, San Diego, La Jolla, United States
- Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States, Jason L Choy Laboratory of Single Molecule Biophysics, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States, QB3 Institute, University of California, Berkeley, Berkeley, United States, Kavli Energy NanoSciences Institute, University of California, Berkeley, Berkeley, United States
- Publication Date:
- Research Org.:
- Univ. of California, Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences and Engineering Division; National Institutes of Health (NIH)
- OSTI Identifier:
- 1180127
- Alternate Identifier(s):
- OSTI ID: 1184781; OSTI ID: 1197950
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Published Article
- Journal Name:
- eLife
- Additional Journal Information:
- Journal Name: eLife Journal Volume: 4; Journal ID: ISSN 2050-084X
- Publisher:
- eLife Sciences Publications, Ltd.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Shin, Jae Yen, Lopez-Garrido, Javier, Lee, Sang-Hyuk, Diaz-Celis, Cesar, Fleming, Tinya, Bustamante, Carlos, and Pogliano, Kit. Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum. United States: N. p., 2015.
Web. doi:10.7554/eLife.06474.
Shin, Jae Yen, Lopez-Garrido, Javier, Lee, Sang-Hyuk, Diaz-Celis, Cesar, Fleming, Tinya, Bustamante, Carlos, & Pogliano, Kit. Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum. United States. https://doi.org/10.7554/eLife.06474
Shin, Jae Yen, Lopez-Garrido, Javier, Lee, Sang-Hyuk, Diaz-Celis, Cesar, Fleming, Tinya, Bustamante, Carlos, and Pogliano, Kit. Thu .
"Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum". United States. https://doi.org/10.7554/eLife.06474.
@article{osti_1180127,
title = {Visualization and functional dissection of coaxial paired SpoIIIE channels across the sporulation septum},
author = {Shin, Jae Yen and Lopez-Garrido, Javier and Lee, Sang-Hyuk and Diaz-Celis, Cesar and Fleming, Tinya and Bustamante, Carlos and Pogliano, Kit},
abstractNote = {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.},
doi = {10.7554/eLife.06474},
journal = {eLife},
number = ,
volume = 4,
place = {United States},
year = {Thu May 07 00:00:00 EDT 2015},
month = {Thu May 07 00:00:00 EDT 2015}
}
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https://doi.org/10.7554/eLife.06474
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