bioRxiv Science⌕ Search

Biology subjects

Gachon, E.

Publications and source records attributed to Gachon, E..

2 recordsLinked to original sources

From flagellar motor behavior to bacterial swimming: defining a reference state for motility dynamics in Magnetospirillum gryphiswaldense

Cell motility is powered by the bacterial flagellar motor, a rotary nanomachine whose activity is dynamically modulated by external stimuli. In Magnetospirillum gryphiswaldense, magnetic and chemical inputs are thought to converge at the level of motility control, yet the mechanisms underlying chemotactic regulation remain poorly understood. Here, we seek to establish a quantitative reference-state of a model strain of magnetotactic bacteria through bacterial flagellar motor dynamics. To achieve this goal, magnetotaxis was kept as a natural factor by keeping the earth's magnetic field as the only source of magnetism. To avoid aerotaxis bias, oxygen gradients were removed by implementing two different constant oxygen conditions: environmental oxygen exposure, and limited oxygen exposure. Together, the tethered-cell bacterial flagellar rotational assay and the free-swimming assay in the absence of external stimuli presented in this paper establish a framework for investigating motor and cellular swimming adaptation to magnetic, aerotactic, and chemical signals. Under reference-state conditions, the bacterial flagellar motor showed a tendency to exhibit log-normal distributions for the time spent in each motor state: runs in different directions (counter-clockwise and clockwise), and pause. A semi-Markov reference-state model was developed to provide a quantitative description of bacterial flagellar motor dynamics. The model revealed that cell magnetic polarity modulates the transition pathways leading to the paused state, whereas oxygen, in the absence of a gradient, primarily regulates residence in each motility state.

biophysics↗

A catch-bond drives stator mechanosensitivity in the Bacterial Flagellar Motor

The bacterial flagellar motor (BFM) is the rotary motor which powers the swimming and swarming of many motile bacteria. The torque is provided by stator units, ion motive force powered ion channels known to assemble and disassemble dynamically in the BFM. This turnover is mechano-sensitive, with the number of engaged units dependent upon the viscous load experienced by the motor through the flagellum. However, the molecular mechanism driving BFM mechano-sensitivity is unknown. Here we directly measure the kinetics of arrival and departure of the stator units in individual wild-type motors via analysis of high-resolution recordings of motor speed, while dynamically varying the load on the motor via external magnetic torque. Obtaining the real-time stator stoichiometry before and after periods of forced motor stall, we measure both the number of active stator units at steady-state as a function of the load and the kinetic association and dissociation rates, by fitting the data to a reversible random sequential adsorption model. Our measurements indicate that BFM mechano-sensing relies on the dissociation rate of the stator units, which decreases with increasing load, while their association rate remains constant. This implies that the lifetime of an active stator unit assembled within the BFM increases when a higher force is applied to its anchoring point in the cell wall, providing strong evidence that a catch-bond mechanism can explain the mechano-sensitivity of the BFM.

biophysics↗