Junhua Yuan

Adaptation at the output of the chemotaxis signalling pathway

In the bacterial chemotaxis network, receptor clusters process input, and flagellar motors generate output. Receptor and motor complexes are coupled by the diffusible protein CheY-P. Receptor output (the steady-state concentration of CheY-P) varies from cell to cell. However, the motor is ultrasensitive, with a narrow operating range of CheY-P concentrations. How the match between receptor output and motor input might be optimized is unclear. Here we show that the motor can shift its operating range by changing its composition. The number of FliM subunits in the C-ring increases in response to a decrement in the concentration of CheY-P, increasing motor sensitivity. This shift in sensitivity explains the slow partial adaptation observed in mutants that lack the receptor methyltransferase and methylesterase and why motors show signal-dependent FliM turnover. Adaptive remodelling is likely to be a common feature in the operation of many molecular machines.

Resurrection of the flagellar rotary motor near zero load

Flagellated bacteria, such as Escherichia coli, are propelled by helical flagellar filaments, each driven at its base by a reversible rotary motor, powered by a transmembrane proton flux. Torque is generated by the interaction of stator proteins, MotA and MotB, with a rotor protein FliG. The physiology of the motor has been studied extensively in the regime of relatively high load and low speed, where it appears to operate close to thermodynamic equilibrium. Here, we describe an assay that allows systematic study of the motor near zero load, where proton translocation and movement of mechanical components are rate limiting. Sixty-nanometer-diameter gold spheres were attached to hooks of cells lacking flagellar filaments, and light scattered from a sphere was monitored at the image plane of a microscope through a small pinhole. Paralyzed motors of cells carrying a motA point mutation were resurrected at 23°C by expression of wild-type MotA, and speeds jumped from zero to a maximum value (≈300 Hz) in one step. Thus, near zero load, the speed of the motor is independent of the number of torque-generating units. Evidently, the units act independently (they do not interfere with one another), and there are no intervals during which a second unit can add to the speed generated by the first (the duty ratio is close to 1).

Parity-Violating Electron Deuteron Scattering and the Proton's Neutral Weak Axial Vector Form Factor

We report on a new measurement of the parity-violating asymmetry in quasielastic electron scattering from the deuteron at backward angles at Q2=0.038 (GeV/c)2. This quantity provides a determination of the neutral weak axial vector form factor of the nucleon, which can potentially receive large electroweak corrections. The measured asymmetry A=-3.51+/-0.57 (stat)+/-0.58 (syst) ppm is consistent with theoretical predictions. We also report on updated results of the previous experiment at Q2=0.091 (GeV/c)2, which are also consistent with theoretical predictions.

Ultrasensitivity of an adaptive bacterial motor

The flagellar motor of Escherichia coli adapts to changes in the steady-state level of the chemotaxis response regulator, CheY-P, by adjusting the number of FliM molecules to which CheY-P binds. Previous measurements of motor ultrasensitivity have been made on cells containing different amounts of CheY-P and, thus, different amounts of FliM in flagellar motors. Here, we designed an experiment to measure the sensitivity of motors containing fixed amounts of FliM, finding Hill coefficients about twice as large as those observed before. This ultrasensitivity provides further insights into the motor switching mechanism and plays important roles in chemotaxis signal amplification and coordination of multiple motors. The Hill coefficients observed here appear to be the highest known for allosteric protein complexes, either biological or synthetic. Extreme motor ultrasensitivity broadens our understanding of mechanisms of allostery and serves as an inspiration for future design of synthetic protein switches.

Switching of the bacterial flagellar motor near zero load.

Flagellated bacteria, such as Escherichia coli, are able to swim up gradients of chemical attractants by modulating the direction of rotation of their flagellar motors, which spin alternately clockwise (CW) and counterclockwise (CCW). Chemotactic behavior has been studied under a variety of conditions, mostly at high loads (at large motor torques). Here, we examine motor switching at low loads. Nano-gold spheres of various sizes were attached to hooks (the flexible coupling at the base of the flagellar filament) of cells lacking flagellar filaments in media containing different concentrations of the viscous agent Ficoll. The speeds and directions of rotation of the spheres were measured. Contrary to the case at high loads, motor switching rates increased appreciably with load. Both the CW-->CCW and CCW-->CW switching rates increased linearly with motor torque. Evidently, the switch senses stator-rotor interactions as well as the CheY-P concentration.

Measurements of ultracold-neutron lifetimes in solid deuterium.

We present the first measurements of the survival time of ultracold neutrons (UCNs) in solid deuterium (SD2). This critical parameter provides a fundamental limitation to the effectiveness of superthermal UCN sources that utilize solid ortho-deuterium as the source material. These measurements are performed utilizing a SD2 source coupled to a spallation source of neutrons, providing a demonstration of UCN production in this geometry and permitting systematic studies of the influence of thermal up-scatter and contamination with para-deuterium on the UCN survival time.

Asymmetry in the clockwise and counterclockwise rotation of the bacterial flagellar motor

Cells of Escherichia coli are able to swim up gradients of chemical attractants by modulating the direction of rotation of their flagellar motors, which spin alternately clockwise (CW) and counterclockwise (CCW). Rotation in either direction has been thought to be symmetric and exhibit the same torques and speeds. The relationship between torque and speed is one of the most important measurable characteristics of the motor, used to distinguish specific mechanisms of motor rotation. Previous measurements of the torque–speed relationship have been made with cells lacking the response regulator CheY that spin their motors exclusively CCW. In this case, the torque declines slightly up to an intermediate speed called the “knee speed” after which it falls rapidly to zero. This result is consistent with a “power-stroke” mechanism for torque generation. Here, we measure the torque–speed relationship for cells that express large amounts of CheY and only spin their motors CW. We find that the torque decreases linearly with speed, a result remarkably different from that for CCW rotation. We obtain similar results for wild-type cells by reexamining data collected in previous work. We speculate that CCW rotation might be optimized for runs, with higher speeds increasing the ability of cells to sense spatial gradients, whereas CW rotation might be optimized for tumbles, where the object is to change cell trajectories. But why a linear torque–speed relationship might be optimum for the latter purpose we do not know.

Thermal and Solvent-Isotope Effects on the Flagellar Rotary Motor near Zero Load

In Escherichia coli, the behavior of the flagellar rotary motor near zero load can be studied by scattering light from nanogold spheres attached to proximal hooks of cells lacking flagellar filaments. We used this method to monitor changes in speed when cells were subjected to changes in temperature or shifted from a medium made with H(2)O to one made with D(2)O. In H(2)O, the speed increased with temperature in a near-exponential manner, with an activation enthalpy of 52 +/- 4 kJ/mol (12.0 +/- 1.0 kcal/mol). In D(2)O, the speed increased in a similar manner, with an activation enthalpy of 50 +/- 4 kJ/mol. The speed in H(2)O was higher than that in D(2)O by a factor of 1.53 +/- 0.14. We performed comparison studies of variations in temperature and solvent isotope, using motors operating at high loads. The variations were small, consistent with previous observations. The implications of these results for proton translocation are discussed.

Measurement of electron backscattering in the energy range of neutron beta decay

We report on the first detailed measurements of electron backscattering from low Z targets at energies up to 124 keV. Both energy and angular distributions of the backscattered electrons are measured and compared with electron transport simulations based on the Geant4 and Penelope Monte Carlo simulation codes. Comparisons are also made with previous, less extensive, measurements and with measurements at lower energies.

Measurement of the neutron β -asymmetry parameter A 0 with ultracold neutrons

We present a detailed report of a measurement of the neutron β-asymmetry parameter A_0, the parity-violating angular correlation between the neutron spin and the decay electron momentum, performed with polarized ultracold neutrons (UCN). UCN were extracted from a pulsed spallation solid deuterium source and polarized via transport through a 7-T magnetic field. The polarized UCN were then transported through an adiabatic-fast-passage spin-flipper field region, prior to storage in a cylindrical decay volume situated within a 1-T 2×2π solenoidal spectrometer. The asymmetry was extracted from measurements of the decay electrons in multiwire proportional chamber and plastic scintillator detector packages located on both ends of the spectrometer. From an analysis of data acquired during runs in 2008 and 2009, we report A_0=−0.11966±0.00089_(−0.00140)^(+0.00123), from which we extract a value for the ratio of the weak axial-vector and vector coupling constants of the nucleon, λ=g_A/g_V=−1.27590±0.00239_(−0.00377)^(+0.00331). Complete details of the analysis are presented.