Light chain 2 is a Tctex-type related axonemal dynein light chain that regulates directional ciliary motility in Trypanosoma brucei

Flagellar motility is essential for the cell morphology, viability, and virulence of pathogenic kinetoplastids, including trypanosomes. Trypanosoma brucei flagella exhibit a bending wave that propagates from the flagellum’s tip to its base, rather than base-to-tip as in other eukaryotes. Thousands of dynein motor proteins coordinate their activity to drive ciliary bending wave propagation. Dynein- associated light and intermediate chains regulate the biophysical mechanisms of axonemal dynein. Tctex- type outer arm dynein light chain 2 (LC2) regulates flagellar bending wave propagation direction, amplitude, and frequency in Chlamydomonas reinhardtii. However, the role of Tctex-type light chains in regulating T. brucei motility is unknown. Here, we used a combination of bioinformatics, in-situ molecular tagging, and immunofluorescence microscopy to identify a Tctex-type light chain in the procyclic form of T. brucei (TbLC2). We knocked down TbLC2 expression using RNAi, rescued the knockdown with eGFP- tagged TbLC2, and quantified TbLC2’s effects on trypanosome cell biology and biophysics. We found that TbLC2 knockdown resulted in kinetoplast mislocalization and the formation of multiple cell clusters in cell culture. We also found that TbLC2 knockdown reduced the directional persistence of trypanosome cell swimming, induced an asymmetric ciliary bending waveform, modulated the bias between the base-to- tip and tip-to-base beating modes, and increased the beating frequency. Together, our findings are consistent with a model of TbLC2 as a down-regulator of axonemal dynein activity that stabilizes the forward tip-to-base beating ciliary waveform characteristic of trypanosome cells. Our work sheds light on axonemal dynein regulation mechanisms that contribute to pathogenic kinetoplastids’ unique tip-to-base ciliary beating nature and how those mechanisms underlie dynein-driven ciliary motility more generally.Author SummaryKinetoplastea is a class of ciliated protists that include parasitic trypanosomes, which cause severe disease in people and livestock in tropical regions across the globe. All trypanosomes, including Trypanosoma brucei, require a cilium to provide propulsive force for directional swimming motility, host immune evasion, and various aspects of their cell cycle. Thus, a functional cilium is essential for the virulence of the parasite.Trypanosome cilia exhibit a unique tip-to-base beating mechanism, different from the base-to-tip beating of most other eukaryotic cilia. Multiple ciliary proteins are involved in the complex biophysical and biochemical mechanisms that underly the trypanosome ciliary beating. These include dynein motor proteins that power the beat, dynein-related light chains that regulate the beat, and many other proteins in the nexin-dynein regulatory complex, in the radial spokes, and associated with the central pair of microtubules, for example.Here, we identify a Tctex-type dynein light chain in T. brucei that we named TbLC2 because it has sequence homology, structural similarity, and ciliary localization like LC2 homologs in other organisms. We demonstrate that TbLC2 has critical dynein regulatory functions, with implications on the unique aspects of trypanosome ciliary beating and cellular swimming motility. Our study represents an additional step toward understanding the functions of the trypanosome ciliary proteome, which could provide novel therapeutic targets against the unique aspects of trypanosome ciliary motility.

Sound absorption based on Micro-perforated panels and Acoustic Black Hole principle

Acoustic black holes (ABHs) have been so far investigated mainly for bending wave ma-nipulation in mechanical structures such as beams or plates. The investigations on ABHs for sound wave manipulation, referred to as Sonic black holes (SBHs) are scarce. Existing SBH structure for sound reduction in air is typically formed by putting a set of rings inside a duct wall with decreasing inner radius according to a power law. As such, the structure is very complex and difficult to be practically realized, which hampers the practical application of SBHs for sound reduction. This study explores the possibilities of achieving SBH effects using other types of structural configurations. In particular, micro-perforated panels are proposed to be introduced into the conventional SBH structure, and the simulation results show that the new formed SBH structure is simpler in configuration in terms of number of rings and more efficient in terms of sound energy trapping and dissipation.

Traveling-wave control of the bending wave in a beam for high quality sound radiation

The bending wave generated by the actuator exciting a panel can be controlled to be in the traveling wave form void the structural resonances, which deteriorates the radiated sound if the panel is used as a speaker. Although such traveling-wave control method (TCM) yields a wider effective frequency range than the modal control method, the requirement of using many actuators is the practical problem yet. If a beam is employed instead of a plate as a panel speaker, the number of actuators can be reduced despite a smaller radiating surface than a plate. This study adopts three actuators for the beam control using TCM. An actuator excites the beam in the middle position, and the two actuators near the two edges are used to suppress the reflected waves from the boundaries. The control result shows that the driving-point mobility of the primary actuator is converted into that of an infinite beam, which means that the boundaries are changed into anechoic ones and the structural resonances are eliminated. Accordingly, the beam radiates a smooth sound spectrum without sharp peaks and troughs related to the resonant responses. Effects of material and dimension in determining the effective frequency range are also explored.

Galactic seismology: the evolving “phase spiral” after the Sagittarius dwarf impact

In 2018, the ESA Gaia satellite discovered a remarkable spiral pattern (“phase spiral”) in the z − Vz phase plane throughout the solar neighbourhood, where z and Vz are the displacement and velocity of a star perpendicular to the Galactic disc. In response to Binney & Schönrich’s analytic model of a disc-crossing satellite to explain the Gaia data, we carry out a high-resolution, N-body simulation (N ≈ 108 particles) of an impulsive mass (2 × 1010 M⊙) that interacts with a cold stellar disc at a single transit point. The disc response is complex since the impulse triggers a superposition of two distinct bisymmetric (m = 2) modes − a density wave and a corrugated bending wave − that wrap up at different rates. Stars in the faster density wave wrap up with time T according to φD(R, T) = (ΩD(R) + Ωo) T where φD describes the spiral pattern and ΩD = Ω(R) − κ(R)/2, where κ is the epicyclic frequency. While the pattern speed Ωo is small, it is non-zero. The slower bending wave wraps up according to ΩB ≈ ΩD/2 producing a corrugated wave. The bunching effect of the density wave triggers the phase spiral as it rolls up and down on the bending wave (“rollercoaster” model). The phase spiral emerges slowly about ΔT ≈ 400 Myr after impact. It appears to be a long-lived, disc-wide phenomenon that continues to evolve over most of the 2 Gyr simulation. Thus, given Sagittarius’ (Sgr) low total mass today (Mtot ∼ 3 × 108 M⊙ within 10 kpc diameter), we believe the phase spiral was excited by the disc-crossing dwarf some 1 − 2 Gyr before the recent transit. For this to be true, Sgr must be losing mass at 0.5-1 dex per orbit loop.