Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase
Abstract
SpoIIIE is a homo-hexameric dsDNA translocase responsible for completing chromosome segregation in Bacillus subtilis . Here, we use a single-molecule approach to monitor SpoIIIE translocation when challenged with neutral-backbone DNA and non-hydrolyzable ATP analogs. We show that SpoIIIE makes multiple essential contacts with phosphates on the 5'→3' strand in the direction of translocation. Using DNA constructs with two neutral-backbone segments separated by a single charged base pair, we deduce that SpoIIIE’s step size is 2 bp. Finally, experiments with non-hydrolyzable ATP analogs suggest that SpoIIIE can operate with non-consecutive inactive subunits. We propose a two-subunit escort translocation mechanism that is strict enough to enable SpoIIIE to track one DNA strand, yet sufficiently compliant to permit the motor to bypass inactive subunits without arrest. We speculate that such a flexible mechanism arose for motors that, like SpoIIIE, constitute functional bottlenecks where the inactivation of even a single motor can be lethal for the cell.
- Authors:
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)
- OSTI Identifier:
- 1233979
- Alternate Identifier(s):
- OSTI ID: 1233980; OSTI ID: 1415944
- Grant/Contract Number:
- AC02-05CH11231; R01GM071552; R01GM032543
- 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
Liu, Ninning, Chistol, Gheorghe, and Bustamante, Carlos. Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase. United States: N. p., 2015.
Web. doi:10.7554/eLife.09224.
Liu, Ninning, Chistol, Gheorghe, & Bustamante, Carlos. Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase. United States. https://doi.org/10.7554/eLife.09224
Liu, Ninning, Chistol, Gheorghe, and Bustamante, Carlos. Fri .
"Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase". United States. https://doi.org/10.7554/eLife.09224.
@article{osti_1233979,
title = {Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase},
author = {Liu, Ninning and Chistol, Gheorghe and Bustamante, Carlos},
abstractNote = {SpoIIIE is a homo-hexameric dsDNA translocase responsible for completing chromosome segregation in Bacillus subtilis . Here, we use a single-molecule approach to monitor SpoIIIE translocation when challenged with neutral-backbone DNA and non-hydrolyzable ATP analogs. We show that SpoIIIE makes multiple essential contacts with phosphates on the 5'→3' strand in the direction of translocation. Using DNA constructs with two neutral-backbone segments separated by a single charged base pair, we deduce that SpoIIIE’s step size is 2 bp. Finally, experiments with non-hydrolyzable ATP analogs suggest that SpoIIIE can operate with non-consecutive inactive subunits. We propose a two-subunit escort translocation mechanism that is strict enough to enable SpoIIIE to track one DNA strand, yet sufficiently compliant to permit the motor to bypass inactive subunits without arrest. We speculate that such a flexible mechanism arose for motors that, like SpoIIIE, constitute functional bottlenecks where the inactivation of even a single motor can be lethal for the cell.},
doi = {10.7554/eLife.09224},
journal = {eLife},
number = ,
volume = 4,
place = {United States},
year = {Fri Oct 09 00:00:00 EDT 2015},
month = {Fri Oct 09 00:00:00 EDT 2015}
}
https://doi.org/10.7554/eLife.09224
Web of Science
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