In situ Investigation of Individual Filament Growth in Conducting Bridge Memristor by Contact Scanning Capacitance Microscopy

We report on the application of Contact Scanning Capacitance Microscopy (CSCM) to trace the growth of an individual Ni filament in a ZrO2(Y) film on a Ni sublayer (together with a conductive Atomic Force Microscope probe composing a nanometer-sized virtual memristor). An increasing of the filament length in the course of electro-forming results in an increasing of the capacitance between the probe and the sample, which can be detected by CSCM technique. This way, the filament growth can be monitored in real time in situ.

Emergence and maintenance of different sized actin filaments in a common pool of building blocks

Actin is one of the key structural components of the eukaryotic cytoskeleton that regulates cellular architecture and mechanical properties. Dynamic regulation of actin filament length and organization is essential for the control of many physiological processes including cell adhesion, motility and division. While previous studies have mostly focused on the mechanisms controlling the mean length of individual actin filaments, it remains poorly understood how distinct actin filament populations in cells maintain different size using the same set of molecular building blocks. Here we develop a theoretical model for the length regulation of multiple actin filaments by nucleation and growth rate modulation by actin binding proteins in a limiting pool of monomers. We first show that spontaneous nucleation of actin filaments naturally leads to heterogeneities in filament length distribution. We then investigate the effects of filament growth inhibition by capping proteins and growth promotion by formin proteins on filament length distribution. We find that filament length heterogeneity can be increased by growth inhibition, whereas growth promoters do not significantly affect length heterogeneities. Interestingly, a competition between filament growth inhibitors and growth promoters can give rise to bimodal filament length distribution as well as a highly heterogeneous length distribution with large statistical dispersion. We quantitatively predict how heterogeneity in actin filament length can be modulated by tuning F-actin nucleation and growth rates in order to create distinct filament subpopulations with different lengths.

Kalanchoe benbothae (K. subg. Fernandesiae; Crassulaceae subfam. Kalanchooideae), a new southern African species endemic to KwaZulu-Natal in the Maputaland-Pondoland Region of Endemism

Since 2016 the number of species of Kalanchoe (Crassulaceae subfam. Kalanchooideae) known from southern Africa has increased by more than 50%, from 13 to 20. We here describe the 21st species known from the subcontinent. As far as could be determined, this new species, K. benbothae, is endemic to northeastern KwaZulu-Natal, the eastern-most province of South Africa, with the entire natural geographical distribution range of the species falling within the Maputaland-Pondoland Region of Endemism. Kalanchoe benbothae fits in K. subg. Fernandesiae, a cluster of species with large, paddle- to soup plate-sized leaves, and dense, elongated, often club-shaped inflorescences. The closest relative of K. benbothae is K. luciae, which was described just over 110 years ago, and was the second representative of K. subg. Fernandesiae to have been described. Kalanchoe benbothae differs from K. luciae mostly in its reproductive morphology (corolla lobe colour and orientation, anther colour and presentation, filament length, pistil shape, and size and shape of the nectar scales).

SMC protein RecN drives translocation and remodelling of RecA filament for homology search

While the molecular repertoire of the homologous recombination pathway is well studied, the search mechanism that enables recombination between distant homologous regions is poorly understood. Here, we follow the dynamics of the recombinase RecA, an essential component of homology search, after induction of a single double-strand break on the Caulobacter chromosome. We find that the RecA-nucleoprotein filament translocates in a directional manner in the cell, undergoing several pole-to-pole traversals, until homology search is complete. Simultaneously, the filament undergoes dynamic remodelling; both translocation and dynamic remodelling are contingent on the action of the SMC protein RecN via its ATPase cycle. We provide a stochastic description of RecN regulated changes in filament length during translocation via modulation of RecA assembly-disassembly. Together, the observed RecN driven RecA dynamics points to a novel optimal search strategy.

Scattered migrating colony formation in the filamentous cyanobacterium, Pseudanabaena sp. NIES-4403

Background Bacteria have been reported to exhibit complicated morphological colony patterns on solid media, depending on intracellular, and extracellular factors such as motility, cell propagation, and cell-cell interaction. We isolated the filamentous cyanobacterium, Pseudanabaena sp. NIES-4403 (Pseudanabaena, hereafter), that forms scattered (discrete) migrating colonies on solid media. While the scattered colony pattern has been observed in some bacterial species, the mechanism underlying such a pattern still remains obscure. Results We studied the morphology of Pseudanabaena migrating collectively and found that this species forms randomly scattered clusters varying in size and further consists of a mixture of comet-like wandering clusters and disk-like rotating clusters. Quantitative analysis of the formation of these wandering and rotating clusters showed that bacterial filaments tend to follow trajectories of previously migrating filaments at velocities that are dependent on filament length. Collisions between filaments occurred without crossing paths, which enhanced their nematic alignments, giving rise to bundle-like colonies. As cells increased and bundles aggregated, comet-like wandering clusters developed. The direction and velocity of the movement of cells in comet-like wandering clusters were highly coordinated. When the wandering clusters entered into a circular orbit, they turned into rotating clusters, maintaining a more stable location. Disk-like rotating clusters may rotate for days, and the speed of cells within a rotating cluster increases from the center to the outmost part of the cluster. Using a mathematical modeling with simplified assumption we reproduced some features of the scattered pattern including migrating clusters. Conclusion Based on these observations, we propose that Pseudanabaena forms scattered migrating colonies that undergo a series of transitions involving several morphological patterns. A simplified model is able to reproduce some features of the observed migrating clusters.

Cross-linkers at growing microtubule ends generate forces that drive actin transport

The actin and microtubule cytoskeletons form active networks in the cell that can contract and remodel, resulting in vital cellular processes as cell division and motility. Motor proteins play an important role in generating the forces required for these processes, but more recently the concept of passive cross-linkers being able to generate forces has emerged. So far, these passive cross-linkers have been studied in the context of separate actin and microtubule systems. Here, we show that cross-linkers also allow actin and microtubules to exert forces on each other. More specifically, we study single actin filaments that are cross-linked to growing microtubule ends, using in vitro reconstitution, computer simulations, and a minimal theoretical model. We show that microtubules can transport actin filaments over large (micrometer-range) distances, and find that this transport results from two antagonistic forces arising from the binding of cross-linkers to the overlap between the actin and microtubule filaments. The cross-linkers attempt to maximize the overlap between the actin and the tip of the growing microtubules, creating an affinity-driven forward condensation force, and simultaneously create a competing friction force along the microtubule lattice. We predict and verify experimentally how the average transport time depends on the actin filament length and the microtubule growth velocity, confirming the competition between a forward condensation force and a backward friction force. In addition, we theoretically predict and experimentally verify that the condensation force is of the order of 0.1pN. Thus, our results reveal a new mechanism for local actin remodelling by growing microtubules.

Multiscale mechanics and temporal evolution of vimentin intermediate filament networks

The cytoskeleton, an intricate network of protein filaments, motor proteins, and cross-linkers, largely determines the mechanical properties of cells. Among the three filamentous components, F-actin, microtubules, and intermediate filaments (IFs), the IF network is by far the most extensible and resilient to stress. We present a multiscale approach to disentangle the three main contributions to vimentin IF network mechanics—single-filament mechanics, filament length, and interactions between filaments—including their temporal evolution. Combining particle tracking, quadruple optical trapping, and computational modeling, we derive quantitative information on the strength and kinetics of filament interactions. Specifically, we find that hydrophobic contributions to network mechanics enter mostly via filament-elongation kinetics, whereas electrostatics have a direct influence on filament–filament interactions.

Formin's nucleation activity influences actin filament length

Formins stimulate actin polymerization by promoting both filament nucleation and elongation. Because nucleation and elongation draw upon a common pool of actin monomers, the rate at which each reaction proceeds influences the other. This interdependent mechanism determines the number of filaments assembled over the course of a polymerization reaction, as well as their equilibrium lengths. In this study, we used kinetic modeling and in vitro polymerization reactions to dissect the contributions of filament nucleation and elongation to the process of formin-mediated actin assembly. We found that the rates of nucleation and elongation evolve over the course of a polymerization reaction. The period over which each process occurs is a key determinant of the total number of filaments that are assembled, as well as their average lengths at equilibrium. Inclusion of formin in polymerization reactions speeds filament nucleation, thus increasing the number and shortening the lengths of filaments that are assembled over the course of the reaction. Although variations in elongation rates produce modest changes in the equilibrium lengths of formin-bound filaments, nucleation constitutes the primary mode of monomer consumption over the course of assembly. Sustained elongation of small numbers of formin-bound filaments therefore requires inhibition of nucleation via monomer sequestration and a low concentration of activated formin. Our results underscore the mechanistic advantage for keeping formin's nucleation efficiency relatively low in cells, where unregulated actin assembly would produce deleterious effects on cytoskeletal dynamics. Under these conditions, differences in the elongation rates mediated by formin isoforms are most likely to impact the kinetics of actin assembly.