First Study of the

New astronomical observations point to a nucleosynthesis picture that goes beyond what was accepted until recently. The intermediate "i" process was proposed as a plausible scenario to explain some of the unusual abundance patterns observed in metal-poor stars. The most important nuclear physics properties entering i-process calculations are the neutron-capture cross sections and they are almost exclusively not known experimentally. Here we provide the first experimental constraints on the ^{139}Ba(n,γ)^{140}Ba reaction rate, which is the dominant source of uncertainty for the production of lanthanum, a key indicator of i-process conditions. This is an important step towards identifying the exact astrophysical site of stars carrying the i-process signature.

Proton Shell Gaps in N=28 Nuclei from the First Complete Spectroscopy Study with FRIB Decay Station Initiator.

The first complete measurement of the ß-decay strength distribution of _{17}^{45}Cl_{28} was performed at the Facility for Rare Isotope Beams (FRIB) with the FRIB Decay Station Initiator during the second FRIB experiment. The measurement involved the detection of neutrons and γ rays in two focal planes of the FRIB Decay Station Initiator in a single experiment for the first time. This enabled an analytical consistency in extracting the ß-decay strength distribution over the large range of excitation energies, including neutron unbound states. We observe a rapid increase in the ß-decay strength distribution above the neutron separation energy in _{18}^{45}Ar_{27}. This was interpreted to be caused by the transitioning of neutrons into protons excited across the Z=20 shell gap. The SDPF-MU interaction with reduced shell gap best reproduced the data. The measurement demonstrates a new approach that is sensitive to the proton shell gap in neutron rich nuclei according to SDPF-MU calculations.

Comprehensive Test of the Brink-Axel Hypothesis in the Energy Region of the Pygmy Dipole Resonance.

The validity of the Brink-Axel hypothesis, which is especially important for numerous astrophysical calculations, is addressed for ^{116,120,124}Sn below the neutron separation energy by means of three independent experimental methods. The γ-ray strength functions (GSFs) extracted from primary γ-decay spectra following charged-particle reactions with the Oslo method and with the shape method demonstrate excellent agreement with those deduced from forward-angle inelastic proton scattering at relativistic beam energies. In addition, the GSFs are shown to be independent of excitation energies and spins of the initial and final states. The results provide a critical test of the generalized Brink-Axel hypothesis in heavy nuclei, demonstrating its applicability in the energy region of the pygmy dipole resonance.

Following the Behavior of the Flagellar Rotary Motor Near Zero Load.

At room temperature at stall, the flagellar motor of the bacterium Escherichia coli exerts a torque of ~1300 pN nm. At zero external load, it spins ~330 Hz. A robust method for studying the motor near zero load is reviewed here.

Restoration of torque in defective flagellar motors.

Paralyzed motors of motA and motB point and deletion mutants of Escherichia coli were repaired by synthesis of wild-type protein. As found earlier with a point mutant of motB, torque was restored in a series of equally spaced steps. The size of the steps was the same for both MotA and MotB. Motors with one torque generator spent more time spinning counterclockwise than did motors with two or more generators. In deletion mutants, stepwise decreases in torque, rare in point mutants, were common. Several cells stopped accelerating after eight steps, suggesting that the maximum complement of torque generators is eight. Each generator appears to contain both MotA and MotB.

Response of the flagellar rotary motor to abrupt changes in extracellular pH.

Cells of a motile Streptococcus were starved, tethered to a quartz coverslip, energized with a potassium diffusion potential, and exposed to sudden decrements in external pH generated by flash photolysis of 2-hydroxyphenyl-1-(2-nitro)phenyl phosphate. The rotation rate of the cells increased following the flash but only after a brief time lag. Lags of the order of 0.1 second were observed in a dilute buffer (0.05 mM), confirming results obtained earlier. These lags were longer when the buffer was prepared in D2O. However, lags as short as 0.01 second were found in more concentrated buffers (1 and 3 mM). In this case, there was no deuterium solvent isotope effect. These differences arise from the extra time required for diffusion of protons from a dilute medium into the cell wall, which has a large buffering capacity. The short lags observed in concentrated media could be inherent to the flagellar motor, but the possibility that they are due to buffered diffusion through the cell wall or to elastic filtering by the tether has not been ruled out.

Mutations in the MotA protein of Escherichia coli reveal domains critical for proton conduction.

The MotA protein of Escherichia coli is an essential component of the torque-generating units that drive the flagellar rotary motor. A variety of evidence indicates that MotA is involved in transmembrane proton conduction. We have now mapped a number of MotA mutants, focusing primarily on those previously shown to be dominant. Fifty-six mutations (all dominant), each causing severe or complete impairment of function, were sequenced and found to correspond to 31 different alleles. All except two of these encoded amino acid substitutions clustered in four hydrophobic, presumably membrane-spanning segments, that together make up only one-third of the length of the polypeptide chain. In contrast, eight mutations (5 dominant), each causing only slight impairment of function (slow alleles), were sequenced and found to specify amino acid substitutions in three hydrophilic domains. The clustering of the mutations provides independent support for the suggestion that MotA is a transmembrane proton channel and places significant constraints on models for the molecular mechanism of ion conduction.

Constraints on flagellar rotation.

The motion of tethered cells of Streptococcus was analyzed at low values of protonmotive force (delta p). Cells repeatedly energized and de-energized stopped at discrete angular positions, indicating a rotational symmetry of barriers to rotation of order 5 or 6. At values of delta p smaller than -30 mV, constraints imposed by these barriers were evident when cells were starved and gradually energized, but not when they were energized first and then gradually de-energized. At values of delta p larger than about -30 mV, the cells behaved as if there were no barriers. Cells spinning in this regime also executed rotational Brownian movement. At energy levels above threshold, the motor determines torque; it does not fix the position of the rotor relative to the stator.

Interacting components of the flagellar motor of Escherichia coli revealed by the two-hybrid system in yeast.

The ability of the flagellar motor of Escherichia coli to switch between clockwise and counterclockwise modes of operation is ultimately responsible for the swimming behavior of the cell. Three motor proteins, FliG, FliM, and FliN, have been implicated in this process. Using the two-hybrid system in Saccharomyces cerevisiae, we demonstrated strong interactions between FliG/FliM,FliM/FliM, and FliM/FliN. A screen for other components that might interact with FliG revealed interactions with FliF (the MS ring protein) and H-NS (a histone-like protein). Regions of proteins important for several of these interactions were identified by mutational analysis. The implications for motor assembly and function are discussed.

A mutant hook-associated protein (HAP3) facilitates torsionally induced transformations of the flagellar filament of Escherichia coli.

Two mutants with defects in hook-associated protein 3 (HAP3) were isolated that exhibit impaired swimming only when they interact with a solid surface or a semisolid matrix. Motility and chemotaxis were normal in liquid media, even in media containing viscous agents, but cells failed to swarm in 0.28% agar. Mutants appeared to carry a full complement of flagella of normal configuration and length. However, filaments rotating counterclockwise close to a glass surface transformed from normal to straight, while filaments rotating clockwise transformed from curly to straight. Both transformations propagated from base to tip, as expected if torsionally induced. The mutations mapped to the middle of flgL, to structural gene for HAP3, and sequence analysis revealed the same coding change in both mutants: a substitution of cysteine for arginine 168. Our results show that the ability of a filament composed of normal flagellin subunits to resist mechanical stress depends on the structure of the protein (HAP3) to which it is attached at its base. The N-terminal sequence of HAP3 was found to be similar to the N-terminal sequence of flagellin, and the possibility that it provides a nucleation site for the C-terminal region of flagellin is discussed.

Torque generation by the flagellar rotary motor.

A review is given of the structure and dynamics of the flagellar rotary motor. Force-generating elements in a motor driving a tethered bacterium (a cell fixed to the substratum by a single flagellum) exert forces of order 20 pN while moving at speeds of order 1 micron/s. Force-generating elements in a motor driving a flagellar filament in a bundle exert forces some 10-fold lower but move at speeds more than 10-fold higher. The motor torque-speed relationship has been measured over a wide dynamic range. Motors strongly resist being driven backwards and are easily broken.