Recent advances and future prospects in bacterial and archaeal locomotion and signal transduction.

Unraveling the structure and function of two-component and chemotactic signaling along with different aspects related to motility of bacteria and archaea are key research areas in modern microbiology. Escherichia coli is the traditional model organism to study chemotaxis signaling and motility. However, the recent study of a wide range of bacteria and even some archaea with different lifestyles has provided new insight into the eco-physiology of chemotaxis, which is essential for the host establishment of different pathogens or beneficial bacteria. The expanded range of model organisms has also permitted the study of chemosensory pathways unrelated to chemotaxis, multiple chemotaxis pathways within an organism, and new types of chemoreceptors. This research has greatly benefitted from technical advances in the field of cryo-microscopy that continues to reveal with increasing resolution the complexity and diversity of large protein complexes like the flagellar motor or chemoreceptor arrays. In addition, sensitive instruments now allow for an increasing number of experiments to be conducted at the single-cell level, thereby revealing information that is beginning to bridge the gap between individual cells and population behavior. Evidence has also accumulated showing that bacteria have evolved different mechanisms for surface sensing, which appears to be mediated by flagella and possibly type IV pili, and that the downstream signaling involves chemosensory pathways and two-component system based processes. Herein we summarize the recent advances and research tendencies in this field as presented at the latest Bacterial Locomotion and Signal Transduction (BLAST XIV) conference.

Bacterial Motility Reveals Unknown Molecular Organization.

The water solubility of lyotropic liquid crystals (LCs) makes them very attractive to study the behavior of biological microorganisms in an environment where local symmetry is broken (as often encountered in nature). Several recent studies have shown a dramatic change in the behavior of flagellated bacteria when swimming in solutions of the lyotropic LC disodium cromoglycate (DSCG). In this study, the movements of Escherichia coli bacteria in DSCG-water solutions of different concentrations are observed to improve our understanding of this phenomenon. In addition, the viscosity of DSCG aqueous solutions is measured as a function of concentration at room temperature. We also experimentally identify a previously undescribed isotropic pretransition zone where bacteria start sticking to each other and to surfaces. Simple estimations show that the unbalanced osmotic pressure induced depletion force might be responsible for this sticking phenomenon. An estimate of the bacteria propulsive force and the DSCG aggregates length (versus concentration) are calculated from the measured viscosity of the medium. All these quantities are found to undergo a strong increase in the pretransition zone, starting at a threshold concentration of 6±1 wt % DSCG that is well below the known isotropic-LC transition (∼10 wt %). This study also shines light on the motility of flagellated bacteria in realistic environments, and it opens new avenues for interesting applications such as the use of motile microorganisms to probe the physical properties of their host or smart bandages that could guide bacteria out of wounds.

World Year of Physics: a direct test of E=mc2.

One of the most striking predictions of Einstein's special theory of relativity is also perhaps the best known formula in all of science: E=mc(2). If this equation were found to be even slightly incorrect, the impact would be enormous--given the degree to which special relativity is woven into the theoretical fabric of modern physics and into everyday applications such as global positioning systems. Here we test this mass-energy relationship directly by combining very accurate measurements of atomic-mass difference, Delta(m), and of gamma-ray wavelengths to determine E, the nuclear binding energy, for isotopes of silicon and sulphur. Einstein's relationship is separately confirmed in two tests, which yield a combined result of 1-Delta(mc2)/E=(-1.4+/-4.4)x10(-7), indicating that it holds to a level of at least 0.00004%. To our knowledge, this is the most precise direct test of the famous equation yet described.

Cyclotron frequency shifts arising from polarization forces.

The cyclotron frequency of a charged particle in a uniform magnetic field B is related to its mass m and charge q by the relationship omega(c) = qB/m. This simple relationship forms the basis for sensitive mass comparisons using ion cyclotron resonance mass spectroscopy, with applications ranging from the identification of biomolecules and the study of chemical reaction rates to determinations of the fine structure constant of atomic spectra. Here we report the observation of a deviation from the cyclotron frequency relationship for polarizable particles: in high-accuracy measurements of a single CO+ ion, a dipole induced in the orbiting ion shifts the measured cyclotron frequency. We use this cyclotron frequency shift to measure non-destructively the quantum state of the CO+ ion. The effect also provides a means to determine to a few per cent the body-frame dipole moment of CO+, thus establishing a method for measuring dipole moments of molecular ions for which few comparably accurate measurements exist. The general perturbation that we describe here affects the most precise mass comparisons attainable today, with applications including direct tests of Einstein's mass-energy relationship and charge-parity-time reversal symmetry, and possibly the weighing of chemical bonds.

Transient locking of the hook procures enhanced motility to flagellated bacteria.

Flagellated bacteria often proliferate in inhomogeneous environments, such as biofilms, swarms and soil. In such media, bacteria are observed to move efficiently only if they can get out of "dead ends" by changing drastically their swimming direction, and even to completely reverse it. Even though these reorientations are ubiquitous, we have only recently begun to describe and understand how they happen. In the present work, we visualized the flagella of bacteria swimming in a soft agar solution. The surprising observation that the filaments do not rotate while being flipped from one side of the cell to the other suggests that reversals are driven directly by the motor rather than by the thrust created by the rotating filament. This was confirmed by observing bacteria in a liquid crystal, where the linear movement of bacteria greatly simplifies the analysis. These observations suggest that the reversal and reorientation processes involve a temporary locking of the flagellum's hook, which is the normally flexible joint between the rotary motor and the long helical filament that propels the cell. This newly described locked-hook mode occurs only when the motor switches to a clockwise rotation. That correlates with other phenomena that are triggered by a switch in one direction and not the other.

Variability in bacterial flagella re-growth patterns after breakage.

Many bacteria swim through liquids or crawl on surfaces by rotating long appendages called flagella. Flagellar filaments are assembled from thousands of subunits that are exported through a narrow secretion channel and polymerize beneath a capping scaffold at the tip of the growing filament. The assembly of a flagellum uses a significant proportion of the biosynthetic capacities of the cell with each filament constituting ~1% of the total cell protein. Here, we addressed a significant question whether a flagellar filament can form a new cap and resume growth after breakage. Re-growth of broken filaments was visualized using sequential 3-color fluorescent labeling of filaments after mechanical shearing. Differential electron microscopy revealed the formation of new cap structures on broken filaments that re-grew. Flagellar filaments are therefore able to re-grow if broken by mechanical shearing forces, which are expected to occur frequently in nature. In contrast, no re-growth was observed on filaments that had been broken using ultrashort laser pulses, a technique allowing for very local damage to individual filaments. We thus conclude that assembly of a new cap at the tip of a broken filament depends on how the filament was broken.

Bacterial flagella grow through an injection-diffusion mechanism.

The bacterial flagellum is a self-assembling nanomachine. The external flagellar filament, several times longer than a bacterial cell body, is made of a few tens of thousands subunits of a single protein: flagellin. A fundamental problem concerns the molecular mechanism of how the flagellum grows outside the cell, where no discernible energy source is available. Here, we monitored the dynamic assembly of individual flagella using in situ labelling and real-time immunostaining of elongating flagellar filaments. We report that the rate of flagellum growth, initially ∼1,700 amino acids per second, decreases with length and that the previously proposed chain mechanism does not contribute to the filament elongation dynamics. Inhibition of the proton motive force-dependent export apparatus revealed a major contribution of substrate injection in driving filament elongation. The combination of experimental and mathematical evidence demonstrates that a simple, injection-diffusion mechanism controls bacterial flagella growth outside the cell.

An ion balance for ultra-high-precision atomic mass measurements.

We have developed the analog of a double-pan balance for determining the masses of single molecular ions from the ratio of their two cyclotron frequencies. By confining two different ions on the same magnetron orbit in a Penning trap, we balance out many sources of noise and error (such as fluctuations of the magnetic field). To minimize the systematic error associated with the Coulomb interaction between the two ions, they are kept about 1 millimeter apart from each other, resulting in fractional uncertainty below 1 x 10(-11). Such precision opens the door to numerous applications of mass spectrometry, including metrology, fundamental physics, and weighing chemical bonds.