DOI: 10.5061/dryad.v1k54
title: Data from: Use of a microfluidic platform to uncover basic features of energy and environmental stress responses in individual cells of Bacillus subtilis

description: Bacteria use a variety of stress-sensing systems to sense and respond to
 diverse stressors and to ensure their survival under adverse conditions.
 The gram-positive bacterium Bacillus subtilis responds to energy stress
 (ATP depletion) and to environmental stressors using two distinct
 stress-sensing pathways that converge on the alternative sigma factor σB
 to provoke a general stress response. Past efforts to study the σB stress
 response in bulk culture and on agarose pads were unable to visualize the
 responses of individual cells under tightly controlled conditions for
 extended periods of time. Here we use a microfluidics-based strategy to
 discern the basic features of σB activation in single cells in response to
 energy and environmental stress, both immediately upon stressor exposure
 and for tens of generations thereafter. Upon energy stress at various
 levels of stressor, cells exhibited fast, transient, and
 amplitude-modulated responses but not frequency modulation as previously
 reported. Upon environmental stress, which is mediated by the stressosome
 complex, wild-type cells primarily exhibited a transient and
 amplitude-modulated response. However, mutant cells producing only one of
 the four paralogous RsbR stressosome proteins showed striking and
 previously unseen differences. Whereas RsbRA-only cells mimicked the wild
 type, RsbRC-only cells displayed a slower but sustained overall response
 composed of repeated activation events in single cells.
description: 1_20160505_WT_40CCCP_tempResponse of wild-type cells to the onset of 40 μM CCCP. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 1.1_20160505_WT_40CCCP.tar2_20160512_WT_25CCCPResponse of wild-type cells to the onset of 25 μM CCCP. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 2.3_20160527_WT_55CCCP_tempResponse of wild-type cells to the onset of 55 μM CCCP. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 3.3_20160527_WT_55CCCP.tar4_20160602_WT_DMSO_tempResponse of wild-type cells to DMSO vehicle. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 4.4_20160602_WT_DMSO.tar5_20160608_del_rsbP_55CCCP_tempResponse of ∆rsbP cells to the onset of 55 μM CCCP. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 5.5_20160608_del_rsbP_55CCCP.tar6_20170612_WT_60MPA_tempResponse of wild-type cells to the onset of 60 μg/ml mycophenolic acid. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 6.6_20170612_WT_60MPA.tar7_20161004_WT_1EtOH_tempResponse of wild-type cells to the onset of 1% ethanol. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 7.7_20161004_WT_1EtOH.tar8_20161012_WT_2EtOH_tempResponse of wild-type cells to the onset of 2% ethanol. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 8.8_20161012_WT_2EtOH.tar9_20170309_DU_2EtOH_tempResponse of ∆rsbU cells to the onset of 2% ethanol. Representative MTC1920 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 9.9_20170309_DU_2EtOH.tar10_20161006_WT_4EtOH_tempResponse of wild-type cells to the onset of 4% ethanol. Representative MTC1801 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 10.10_20161006_WT_4EtOH.tar11_20161020_RA_2EtOH_tempResponse of RsbRA-only cells to the onset of 2% ethanol. Representative MTC1761 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 11.11_20161020_RA_2EtOH.tar12_20160819_RC_2EtOH_tempResponse of RsbRC-only cells to the onset of 2% ethanol. Representative MTC1765 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 12.12_20160819_RC_2EtOH.tar13_20170104_RB_2EtOH_tempResponse of RsbRB-only cells to the onset of 2% ethanol. Representative MTC1763 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 13.13_20170104_RB_2EtOH.tar14_20161117_RD_2EtOH_tempResponse of RsbRD-only cells to the onset of 2% ethanol. Representative MTC1767 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 14.14_20161117_RD_2EtOH.tar15_20161026_RA_1EtOH_tempResponse of RsbRA-only cells to the onset of 1% ethanol. Representative MTC1761 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 15.15_20161026_RA_1EtOH.tar16_20160921_RC_1EtOH_tempResponse of RsbRC-only cells to the onset of 1% ethanol. Representative MTC1765 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 16.16_20160921_RC_1EtOH.tar17_20160915_RC_4EtOH_tempResponse of RsbRC-only cells to the onset of 4% ethanol. Representative MTC1765 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 17.17_20160915_RC_4EtOH.tar18_20161017_RA_4EtOH_tempResponse of RsbRA-only cells to the onset of 4% ethanol. Representative MTC1761 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. This experiment corresponds to Movie 18.18_20161017_RA_4EtOH.tar19_20160822_DUP_LB_tempResponse of ∆rsbP ∆rsbU cells. Representative MTC1930 cell lineages are shown, with the mother cells oriented toward the bottom of the frame and the feeding/waste channel toward the top of the frame. The cells are kept in LB with no medium switch to assess whether there is any σB activation in the absence of both known activating phosphatases. This experiment corresponds to Figure S10.19_20160822_DUP_LB.tar