Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

doi:10.7554/eLife.09224

Title: Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

Journal Article · Fri Oct 09 00:00:00 EDT 2015 · eLife

DOI:https://doi.org/10.7554/eLife.09224· OSTI ID:1233979

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)

Grant/Contract Number:: AC02-05CH11231; R01GM071552; R01GM032543

OSTI ID:: 1233979

Alternate ID(s):: OSTI ID: 1233980; OSTI ID: 1415944

Journal Information:: eLife, Journal Name: eLife Vol. 4; ISSN 2050-084X

Publisher:: eLife Sciences Publications, Ltd.Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 12 works

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