Publications

Highly redundant pathways often contain components whose functions are difficult to decipher from the responses induced by genetic or molecular perturbations. Here, we present a statistical approach that samples and registers events observed in images of intrinsic fluctuations in unperturbed cells to establish the functional hierarchy of events in systems with redundant pathways. We apply this approach to study the recruitment of actin assembly factors involved in the protrusion of the cell membrane. We find that the formin mDia1, along with nascent adhesion components, is recruited to the leading edge of the cell before protrusion onset, initiating linear growth of the lamellipodial network. Recruitment of Arp2/3, VASP, cofilin, and the formin mDia2 then promotes sustained exponential growth of the network. Experiments changing membrane tension suggest that Arp2/3 recruitment is mechano-responsive. These results indicate that cells adjust the overlapping contributions of multiple factors to actin filament assembly during protrusion on a ten-second timescale and in response to mechanical cues.

Filopodia are finger-like structures containing parallel bundles of actin filaments that are central to eukaryotic cell motility in a variety of contexts. K. Lee et al. (p. 1341) reconstituted filopodia-like structures that grow from supported lipid bilayers to explore filopodia assembly. A structural transition from actin networks to parallel bundles was observed that mediated self-assembly of filopodia-tip complexes on the membranes. Actin bundle structures formed on lipid bilayers give insight into formation of the finger-like structures involved in cell migration. Filopodia are finger-like protrusive structures, containing actin bundles. By incubating frog egg extracts with supported lipid bilayers containing phosphatidylinositol 4,5 bisphosphate, we have reconstituted the assembly of filopodia-like structures (FLSs). The actin assembles into parallel bundles, and known filopodial components localize to the tip and shaft. The filopodia tip complexes self-organize—they are not templated by preexisting membrane microdomains. The F-BAR domain protein toca-1 recruits N-WASP, followed by the Arp2/3 complex and actin. Elongation proteins, Diaphanous-related formin, VASP, and fascin are recruited subsequently. Although the Arp2/3 complex is required for FLS initiation, it is not essential for elongation, which involves formins. We propose that filopodia form via clustering of Arp2/3 complex activators, self-assembly of filopodial tip complexes on the membrane, and outgrowth of actin bundles.

We present a statistical physics model to describe the stochastic behaviorof ion transport and channel transitions under an applied membrane voltage.To get pertinent ideas we apply our general theoretical scheme to ananalytically tractable model of the channel with a deep binding site whichinteracts with the permeant ions electrostatically. It is found that theinteraction is modulated by the average ionic occupancy in the bindingsite, which is enhanced by the membrane voltage increases. Above acritical voltage, the interaction gives rise to a emergence of a newconducting state along with shift of S4 charge residues in the channel.This exploratory study calls for further investigations to correlate thecomplex transition behaviors with a variety of ion channels, withparameters in the model, potential energy parameters, voltage, and ionic concentration.

An ion channel is a macromolecular machinery (NANOMACHINE) which regulates the ionic conduction through biomembranes. The ion channels are fundamental in every thought, every perception, every movement, and every heartbeat. The relevant biological dynamics in mesoscopic level is the overdamped slow dynamics. We present a stochastic model to describe the coupled behaviors of ion transport and channel conformation under an applied (trans)membrane potential. We apply our general theoretical formulae to an analytically tractable model of channel with a deep binding site which interacts with the permeant ions electrostatically or entropically. The interaction is found to be modulated by the ionic occupancy which is enhanced by the membrane potential. Above a critical interaction strength or a membrane potential, the interaction gives rise to an emergence of a new conductance state, via a channel conformational transition. This is the self-organization generic to strongly coupled stochastic processes.

We investigate the effects of nonequilibrium fluctuations on ionic transport through ion channels in membranes using the concept of localized ratchet. Due to the localization, the ionic population in the binding site can be enhanced or suppressed depending upon ionic potential and its fluctuations, affecting the gating kinetics of the channel. The localized dichotomic fluctuations of ionic potential are shown to give rise to a current reversal differing from the results of periodic ratchets. It is also found that strong correlations between binding energy and membrane potential fluctuations induce resonancelike behaviors in ionic current as the fluctuating rate varies.