Molecular Motors Research Articles

Stochastic nucleosome disassembly mediated by remodelers and histone fragmentation.

We construct and analyze monomeric and multimeric models of the stochastic disassembly of a single nucleosome. Our monomeric model predicts the time needed for a number of histone-DNA contacts to spontaneously break, leading to dissociation of a non-fragmented histone from DNA. The dissociation process can be facilitated by DNA binding proteins or processing molecular motors that compete with histones for histone-DNA contact sites. Eigenvalue analysis of the corresponding master equation allows us to evaluate histone detachment times under both spontaneous detachment and protein-facilitated processes. We find that competitive DNA binding of remodeling proteins can significantly reduce the typical detachment time but only if these remodelers have DNA-binding affinities comparable to those of histone-DNA contact sites. In the presence of processive motors, the histone detachment rate is shown to be proportional to the product of the histone single-bond dissociation constant and the speed of motor protein procession. Our simple intact-histone model is then extended to allow for multimeric nucleosome kinetics that reveal additional pathways of disassembly. In addition to a dependence of complete disassembly times on subunit-DNA contact energies, we show how histone subunit concentrations in bulk solutions can mediate the disassembly process by rescuing partially disassembled nucleosomes. Moreover, our kinetic model predicts that remodeler binding can also bias certain pathways of nucleosome disassembly, with higher remodeler binding rates favoring intact-histone detachment.

Conformational selection accelerates catalysis by an organocatalytic molecular motor

Conformational selection accelerates catalysis by an organocatalytic molecular motor

Selection for Size in Molecular Self-Assembly Drives the DeNovo Evolution of a Molecular Machine.

The functioning of machines typically requires a concerted action of their parts. This requirement also holds for molecular motors that drive vital cellular processes and imposes constraints on their conformational changes as well as the rates at which they occur. It remains unclear whether, during evolution, features required for functional molecular machines can emerge simultaneously or require sequential adaptation to different selection pressures. We address this question by theoretically analyzing the evolution of filament treadmilling. This process refers to the self-assembly of linear polymers that grow and shrink at equal rates at their opposite ends. It constitutes a simple biological molecular machine that is involved in bacterial cell division and requires that several conditions are met. In our simulation framework, treadmilling emerges as a consequence of selecting for a target average polymer length. We discuss why other forms of assembly dynamics, which also reach the imposed target length, do not emerge in our simulations. Our work shows that complex molecular functions can evolve denovo under selection for a single physical feature.

Nonequilibrium steady states in coupled asymmetric and symmetric exclusion processes.

We propose and study a one-dimensional (1D) model consisting of two lanes with open boundaries. One of the lanes executes diffusive and the other lane driven unidirectional or asymmetric exclusion dynamics, which are mutually coupled through particle exchanges in the bulk. We elucidate the generic nonuniform steady states in this model. We show that in a parameter regime, where hopping along the TASEP lane, diffusion along the SEP lane, and the exchange of particles between the TASEP and SEP lanes compete, the SEP diffusivity D appears as a tuning parameter for both the SEP and TASEP densities for a given exchange rate in the nonequilibrium steady states of this model. Indeed, D can be tuned to achieve phase coexistence in the asymmetric exclusion dynamics together with spatially smoothly varying density in the diffusive dynamics in the steady state. We obtain phase diagrams of the model using mean field theories, and corroborate and complement the results with stochastic Monte Carlo simulations. This model reduces to an isolated open totally asymmetric exclusion process (TASEP) and an open TASEP with bulk particle nonconserving Langmuir kinetics (LK), respectively, in the limits of vanishing and diverging particle diffusivity in the lane executing diffusive dynamics. Thus, this model works as an overarching general model, connecting both pure TASEPs and TASEPs with LK in different asymptotic limits. We further define phases in the SEP and obtain phase diagrams and show their correspondence with the TASEP phases. In addition to its significance as a 1D driven, diffusive model, this model also serves as a simple reduced model for cell biological transport by molecular motors undergoing diffusive and directed motion inside eukaryotic cells.

Regular Arrays of Rod-Shaped Molecular Photoswitches: Synthesis, Preparation, Characterization, and Selective Photoswitching within Mono- and Bilayer Systems.

We assembled photoresponsive mono- and bilayer systems with well-defined properties from rod-shaped molecules equipped with different photoswitches. Using properly chosen chromophores (diarylethene-based switch and unidirectional light-driven molecular motor), we then selectively targeted layers made of the same types of photoswitches using appropriate monochromatic light. UV-vis analysis confirmed smooth and unrestricted photoisomerization. To achieve this, we synthesized a new class of triptycene-based molecular pedestals adept at forming sturdy Langmuir-Blodgett films on a water-air interface. The films were smoothly transferred to gold and quartz surfaces. Repeated deposition afforded bilayer systems: one layer containing diarylethene-based photoswitches and the other a unidirectional light-driven molecular motor. Structural analysis of both mono- and bilayer systems revealed the molecules to be tilted with carboxylic functions pointing to the surface. At least two different polymorphs differing in monolayer thickness and tilt angle (~40° and ~60°) were identified on the gold surface.

Emergence of diverse patterns driven by molecular motors in the motility assay.

Actomyosin contractility originating from interactions between F-actin and myosin motors in the actin cytoskeleton generates mechanical forces and drives a wide range of cellular processes including cell migration and cytokinesis. To probe the interactions between F-actin and myosin motors, the myosin motility assay has been popularly employed, which consists of myosin heads attached to a glass surface and F-actins gliding on the surface via interactions with the heads. Several experiments have shown that F-actins move in a collective fashion due to volume-exclusion effects between neighboring F-actins. Furthermore, Computational models have shown how changes in key parameters lead to diverse pattern formation in motility assay. However, in most of the computational models, myosin motors were implicitly considered by applying a constant propulsion force to filaments to reduce computational cost. This simplification limits the physiological relevance of the insights provided by the models and potentially leads to artifacts. In this study, we employed an agent-based computational model for the motility assay with explicit immobile motors interacting with filaments. We rigorously account for the kinetics of myosin motors including the force-velocity relationship for walking and the binding and unbinding behaviors. We probed the effects of the length, rigidity, and concentration of filaments and repulsive strength on collective movements and pattern formation. It was found that four distinct types of structures-homogeneous networks, flocks, bands, and rings-emerged as a result of collisions between gliding filaments. We further analyzed the frequency and morphology of these structures and the curvature, alignment, and rotational motions of filaments. Our study provides better insights into the origin and properties of patterns formed by gliding filaments beyond what was shown before.

Acidification of the phagosome orchestrates the motor forces directing its transport

Phagosomes are dynamic organelles formed by macrophages to capture and destroy microbial pathogens. Phagosome transport from the cell periphery to the perinuclear region, is essential for fusion with lysosomes and the elimination of pathogens. Molecular motors, kinesin and dynein, generate opposing forces, transporting the phagosome away from and towards the lysosome, respectively. Luminal acidification plays a crucial role in determining the net directional movement of the phagosome. The mechanics of this regulation are not known. In this study, we used the sodium proton exchanger NHE9 to selectively modulate phagosomal acidification in macrophages. We then investigated its impact on the mechanical properties of kinesin and dynein motors through optical trapping experiments. We observed a negative correlation between the tenacity of dynein motors and pH under high resistive forces. Reduced luminal acidification impaired generation of dynein cooperative forces, which are crucial for transporting the phagosome to the lysosome. Conversely, the kinesin-powered motility of phagosomes is enabled by a decrease in phagosomal acidification. Given the various methods pathogens employ to limit phagosomal acidification, our findings are highly significant in the context of host-pathogen interactions.

How do molecular motors fold the genome?

A potential mechanism of DNA loop extrusion by molecular motors is discussed.

Coordination of Pickpocket ion channel delivery and dendrite growth in Drosophila sensory neurons.

Sensory neurons enable an organism to perceive external stimuli, which is essential for survival. The sensory capacity of a neuron depends on the elaboration of its dendritic arbor and the localization of sensory ion channels to the dendritic membrane. However, it is not well understood when and how ion channels localize to growing sensory dendrites and whether their delivery is coordinated with growth of the dendritic arbor. We investigated the localization of the DEG/ENaC/ASIC ion channel Pickpocket (Ppk) in the peripheral sensory neurons of developing fruit flies. We used CRISPR-Cas9 genome engineering approaches to tag endogenous Ppk1 and visualize it live, including monitoring Ppk1 membrane localization via a novel secreted split-GFP approach. Fluorescently tagged endogenous Ppk1 localizes to dendrites, as previously reported, and, unexpectedly, to axons and axon terminals. In dendrites, Ppk1 is present throughout actively growing dendrite branches and is stably integrated into the neuronal cell membrane during the expansive growth of the arbor. Although Ppk channels are dispensable for dendrite growth, we found that an over-active channel mutant severely reduces dendrite growth, likely by acting at an internal membrane and not the dendritic membrane. Our data reveal that the molecular motor dynein and recycling endosome GTPase Rab11 are needed for the proper trafficking of Ppk1 to dendrites. Based on our data, we propose that Ppk channel transport is coordinated with dendrite morphogenesis, which ensures proper ion channel density and distribution in sensory dendrites.

Recently recycled synaptic vesicles use multi-cytoskeletal transport and differential presynaptic capture probability to establish a retrograde net flux during ISVE in central neurons.

Presynapses locally recycle synaptic vesicles to efficiently communicate information. During use and recycling, proteins on the surface of synaptic vesicles break down and become less efficient. In order to maintain efficient presynaptic function and accommodate protein breakdown, new proteins are regularly produced in the soma and trafficked to presynaptic locations where they replace older protein-carrying vesicles. Maintaining a balance of new proteins and older proteins is thus essential for presynaptic maintenance and plasticity. While protein production and turnover have been extensively studied, it is still unclear how older synaptic vesicles are trafficked back to the soma for recycling in order to maintain balance. In the present study, we use a combination of fluorescence microscopy, hippocampal cell cultures, and computational analyses to determine the mechanisms that mediate older synaptic vesicle trafficking back to the soma. We show that synaptic vesicles, which have recently undergone exocytosis, can differentially utilize either the microtubule or the actin cytoskeleton networks. We show that axonally trafficked vesicles traveling with higher speeds utilize the microtubule network and are less likely to be captured by presynapses, while slower vesicles utilize the actin network and are more likely to be captured by presynapses. We also show that retrograde-driven vesicles are less likely to be captured by a neighboring presynapse than anterograde-driven vesicles. We show that the loss of synaptic vesicle with bound molecular motor myosin V is the mechanism that differentiates whether vesicles will utilize the microtubule or actin networks. Finally, we present a theoretical framework of how our experimentally observed retrograde vesicle trafficking bias maintains the balance with previously observed rates of new vesicle trafficking from the soma.

Excited-State Dynamics Simulations of a Light-Driven Molecular Motor in Solution.

Molecular motors, where light can be transformed into motion, are promising in the design of nanomechanical devices. For applications, however, finding relationships between molecular motion and the environment is important. Here, we report the study of excited-state dynamics of an overcrowded alkene in solution using a hybrid quantum mechanics/molecular mechanics (QM/MM) approach combined with excited-state molecular dynamics simulations. Using QM/MM surface-hopping trajectories, we calculated time-resolved emission and transient absorption spectra. These show the rise of a short-lived Franck-Condon state, followed by the formation of a dark state in the first 150 fs before the molecular motor relaxes to the ground state in about 1 ps. From the analysis of radial distribution functions, we infer that the orientation of the solvent with respect to the molecular motor in the electronic excited state is similar to that in the ground state during the photoisomerization.

On the use of thermal forces to probe kinesin's response to force.

The stepping dynamics of cytoskeletal motor proteins determines the dynamics of cargo transport. In its native cellular environment, a molecular motor is subject to forces from several sources including thermal forces and forces ensuing from the interaction with other motors bound to the same cargo. Understanding how the individual motors respond to these forces can allow us to predict how they move their cargo when part of a team. Here, using simulation, we show that details of how the kinesin motor responds to small assisting forces-which, at the moment, are not experimentally constrained-can lead to significant changes in cargo dynamics. Using different models of the force-dependent detachment probability of the kinesin motor leads to different predictions on the run-length of the cargo they carry. These differences emerge from the thermal forces acting on the cargo and transmitted to the motor through the motor tail that tethers the motor head to the microtubule. We show that these differences appear for cargo carried by individual motors or motor teams, and use our findings to propose the use of thermal forces as a probe of kinesin's response to force in this otherwise inaccessible force regime.

Local glycolysis fuels actomyosin contraction during axonal retraction.

In response to repulsive cues, axonal growth cones can quickly retract. This requires the prompt activity of contractile actomyosin, which is formed by the non-muscle myosin II (NMII) bound to actin filaments. NMII is a molecular motor that provides the necessary mechanical force at the expense of ATP. Here, we report that this process is energetically coupled to glycolysis and is independent of cellular ATP levels. Induction of axonal retraction requires simultaneous generation of ATP by glycolysis, as shown by chemical inhibition and genetic knock-down of GAPDH. Co-immunoprecipitation and proximal-ligation assay showed that actomyosin associates with ATP-generating glycolytic enzymes and that this association is strongly enhanced during retraction. Using microfluidics, we confirmed that the energetic coupling between glycolysis and actomyosin necessary for axonal retraction is localized to the growth cone and near axonal shaft. These results indicate a tight coupling between on-demand energy production by glycolysis and energy consumption by actomyosin contraction suggesting a function of glycolysis in axonal guidance.

Artificial molecular machines: Design and observation

AbstractNatural molecular machines have inspired the development of artificial molecular machines, which have the potential to revolutionize several areas of technology. Artificial molecular machines commonly employ molecular switches, molecular motors, and molecular shuttles as fundamental building blocks. The observation of artificial molecular machines constructed by these building blocks can be highly challenging due to their small sizes and intricate behaviors. The use of modern instrumentation and advanced observational techniques plays a crucial role in the observation and characterization of molecular machines. Furthermore, a well‐designed molecular structure is also a critical factor in making molecular machines more observable. This review summarizes the common methods from diverse perspectives used to observe molecular machines and emphasizes the significance of comprehending their behaviors in the design of superior artificial molecular machines.

Spatacsin regulates directionality of lysosome trafficking by promoting the degradation of its partner AP5Z1.

The endoplasmic reticulum (ER) forms contacts with the lysosomal compartment, regulating lysosome positioning and motility. The movements of lysosomes are controlled by the attachment of molecular motors to their surface. However, the molecular mechanisms by which ER controls lysosome dynamics are still elusive. Here, using mouse brain extracts and mouse embryonic fibroblasts, we demonstrate that spatacsin is an ER-resident protein regulating the formation of tubular lysosomes, which are highly dynamic. Screening for spatacsin partners required for tubular lysosome formation showed spatacsin to act by regulating protein degradation. We demonstrate that spatacsin promotes the degradation of its partner AP5Z1, which regulates the relative amount of spastizin and AP5Z1 at lysosomes. Spastizin and AP5Z1 contribute to regulate tubular lysosome formation, as well as their trafficking by interacting with anterograde and retrograde motor proteins, kinesin KIF13A and dynein/dynactin subunit p150Glued, respectively. Ultimately, investigations in polarized mouse cortical neurons in culture demonstrated that spatacsin-regulated degradation of AP5Z1 controls the directionality of lysosomes trafficking. Collectively, our results identify spatacsin as a protein regulating the directionality of lysosome trafficking.

Control of Osmotic-Engine-Driven Liposomes Using Biological Nanopores

Liposome-based molecular robots that molecular systems are integrated into a giant liposome have been proposed; they are expected to be applied in the fields of medicine, environmental science, food science, and energy science. However, the performance of these molecular robotic components, including intelligence, sensors, and actuators, still hinders their practical use. In particular, the actuators used in the molecular robots, such as molecular motors, do not provide sufficient performance to move the giant liposomes. Hence, we propose an osmotic-engine-driven liposome and demonstrate the migration of liposomes in a microfluidic channel by applying a salt concentration difference between the front and rear of the liposome. Although the migration mechanism is simple and has the potential to provide sufficient mobility performance, control techniques for the movement speed and on/off switching are not established. Herein, we describe a speed control method of osmotic-engine-driven liposomes using pore-forming membrane proteins. In this study, we evaluated the effect of reconstituted α-hemolysin (αHL) nanopores on the water permeability through lipid bilayers. Thereafter, we demonstrated the change in displacement speeds of liposomes with and without nanopores. We expect the speed control method using nanopores to be applied to the liposome-based molecular robots.

A Chemically Inducible Organelle Rerouting Assay to Probe Primary Cilium Assembly, Maintenance, and Disassembly in Cultured Cells.

The primary cilium is a conserved, microtubule-based organelle that protrudes from the surface of most vertebrate cells as well as sensory cells of many organisms. It transduces extracellular chemical and mechanical cues to regulate diverse cellular processes during development and physiology. Loss-of-function studies via RNA interference and CRISPR/Cas9-mediated gene knockouts have been the main tool for elucidating the functions of proteins, protein complexes, and organelles implicated in cilium biology. However, these methods are limited in studying acute spatiotemporal functions of proteins as well as the connection between their cellular positioning and functions. A powerful approach based on inducible recruitment of plus or minus end-directed molecular motors to the protein of interest enables fast and precise control of protein activity in time and in space. In this chapter, we present a chemically inducible heterodimerization method for functional perturbation of centriolar satellites, an emerging membrane-less organelle involved in cilium biogenesis and function. The method we present is based on rerouting of centriolar satellites to the cell center or the periphery in mammalian epithelial cells. We also describe how this method can be applied to study the temporal functions of centriolar satellites during primary cilium assembly, maintenance, and disassembly.

A rhythmically pulsing leaf-spring DNA-origami nanoengine that drives a passive follower

Molecular engineering seeks to create functional entities for modular use in the bottom-up design of nanoassemblies that can perform complex tasks. Such systems require fuel-consuming nanomotors that can actively drive downstream passive followers. Most artificial molecular motors are driven by Brownian motion, in which, with few exceptions, the generated forces are non-directed and insufficient for efficient transfer to passive second-level components. Consequently, efficient chemical-fuel-driven nanoscale driver–follower systems have not yet been realized. Here we present a DNA nanomachine (70 nm × 70 nm × 12 nm) driven by the chemical energy of DNA-templated RNA-transcription-consuming nucleoside triphosphates as fuel to generate a rhythmic pulsating motion of two rigid DNA-origami arms. Furthermore, we demonstrate actuation control and the simple coupling of the active nanomachine with a passive follower, to which it then transmits its motion, forming a true driver–follower pair.

The Tubulin Code, from Molecules to Health and Disease.

Microtubules are essential dynamic polymers composed of α/β-tubulin heterodimers. They support intracellular trafficking, cell division, cellular motility, and other essential cellular processes. In many species, both α-tubulin and β-tubulin are encoded by multiple genes with distinct expression profiles and functionality. Microtubules are further diversified through abundant posttranslational modifications, which are added and removed by a suite of enzymes to form complex, stereotyped cellular arrays. The genetic and chemical diversity of tubulin constitute a tubulin code that regulates intrinsic microtubule properties and is read by cellular effectors, such as molecular motors and microtubule-associated proteins, to provide spatial and temporal specificity to microtubules in cells. In this review, we synthesize the rapidly expanding tubulin code literature and highlight limitations and opportunities for the field. As complex microtubule arrays underlie essential physiological processes, a better understanding of how cells employ the tubulin code has important implications for human disease ranging from cancer to neurological disorders.

Chasing the rainbow: Exploiting photosensitizers to drive photoisomerization reactions

AbstractPhotoswitchable molecules have garnered considerable attention for their versatility and diverse applications, spanning from solar energy harvesting and storage to drug delivery and molecular motors. The chemical conversions that make photoswitches a desirable system are driven by specific wavelengths of light, which often demand intricate molecular modifications. An alternative approach to achieve the photoisomerization reaction is through energy transfer with photosensitizers. Photosensitizers play a pivotal role in various light‐induced processes and have demonstrated successful applications in photodynamic therapy, dye‐sensitized solar cells, and activating photochemical reactions. Therefore, combining photoswitching systems with sensitizers presents an attractive alternative for advancing light‐responsive material design and enabling innovative light‐controlled technologies. This review summarizes the energy transfer mechanisms and strategies involved in sensitized molecular photoswitchable systems, emphasizing the performance of various combined systems, and potential applications. Furthermore, recent advances and emerging trends in this field are also discussed, offering insights into prospective future directions for the development of light‐responsive materials.