Mesoscopic biology

In this paper we present a qualitative outlook of mesoscopic biology where the typical length scale is of the order of nanometers and the energy scales comparable to thermal energy. Novel biomolecular machines, governed by coded information at the level of DNA and proteins, operate at these length scales in biological systems. In recent years advances in technology have led to the study of some of the design principles of these machines; in particular at the level of an individual molecule. For example, the forces that operate in molecular interactions, the stochasticity involved in these interactions and their spatio-temporal dynamics are beginning to be explored. Understanding such design principles is opening new possibilities in mesoscopic physics with potential applications

Probing collective dynamics of active particles using modulation force spectroscopy

In this letter, we report a method of measuring the dynamic viscosity of self-propelled active particles using an intensity-modulated optical tweezer. We have used a 6 μm trapped polystyrene bead suspended in a bath of motile bacterial cells as a probe. The response function amplitude of the oscillatory bead directly measures the dynamics of the spatiotemporal structure of the motile particles. We find that unlike passive systems, the viscosity is defined by distributions of response function amplitudes that represent the long-range active temporal structures. Appropriate Langevin equations are set up that capture all these essential features

Nanomechanics of membrane tubulation and DNA assembly

We report an interesting regime of tubule formation in multilamellar membrane vesicles. An optically trapped bead is used to apply a localized subpicoNewton force on a cationic vesicle to form a membrane tubule. The force extension curves reveal a saturation phase, with the tubule length extending up to tens of microns, beyond a threshold force 0.6±0.2 pN. We then use the tubule as a sensor for monitoring the dynamics of charge induced DNA integration on cationic membrane vesicles. Our results may also have applications in the development of nanowires and nanofluidic devices

Possible observation of coulomb blockade at room temperature

We have studied the (I-V) characteristics of the tunnel junction formed between the tip and the substrate in an STM at room temperature. We find that in such an arrangement it may be possible to get a junction capacitance ⋍10−19 F and junction conductance < 1 μs. When the junction conductance is < 1 μs strong nonlinearity is observed in the (I-V) characteristics. We explain this nonlinearity as onset of coulomb blockade of tunneling electrons

Maximal fluctuations of confined actomyosin gels: dynamics of the cell nucleus

We investigate the effect of stress fluctuations on the stochastic dynamics of an inclusion embedded in a viscous gel. We show that, in non-equilibrium systems, stress fluctuations give rise to an effective attraction towards the boundaries of the confining domain, which is reminiscent of an active Casimir effect. We apply this generic result to the dynamics of deformations of the cell nucleus and we demonstrate the appearance of a fluctuation maximum at a critical level of activity, in agreement with recent experiments [E. Makhija, D. S. Jokhun, and G. V. Shivashankar, Proc. Natl. Acad. Sci. U.S.A. 113, E32 (2016)].Comment: 12 pages, 5 figure

Soft inclusion in a confined fluctuating active gel

We study stochastic dynamics of a point and extended inclusion within a one dimensional confined active viscoelastic gel. We show that the dynamics of a point inclusion can be described by a Langevin equation with a confining potential and multiplicative noise. Using a systematic adiabatic elimination over the fast variables, we arrive at an overdamped equation with a proper definition of the multiplicative noise. To highlight various features and to appeal to different biological contexts, we treat the inclusion in turn as a rigid extended element, an elastic element and a viscoelastic (Kelvin-Voigt) element. The dynamics for the shape and position of the extended inclusion can be described by coupled Langevin equations. Deriving exact expressions for the corresponding steady state probability distributions, we find that the active noise induces an attraction to the edges of the confining domain. In the presence of a competing centering force, we find that the shape of the probability distribution exhibits a sharp transition upon varying the amplitude of the active noise. Our results could help understanding the positioning and deformability of biological inclusions, eg. organelles in cells, or nucleus and cells within tissues.Comment: 16 pages, 9 figure

Role of actin dependent nuclear deformation in regulating early gene expression

The nucleus of a living cell is constantly undergoing changes in shape and size as a result of various mechanical forces in physiology. These changes correlate with alterations in gene expression, however it is unclear whether nuclear deformation alone is sufficient to elicit these alterations. We used T-cell activation as a model system to test the coupling between nuclear deformation (elongation) and gene expression. Naïve T-cell activation with surrogate antigens resulted in actin dependent nuclear elongation. This was accompanied with Erk and NF-κB signaling to the nucleus to induce CD69 expression. Importantly, inhibiting actin polymerization abolished both nuclear elongation and CD69 expression, while inhibiting Erk, NF-κB or microtubule depolymerization only abolished expression but not elongation. Immobilization of antigen-coated beads, under conditions where actin polymerization was inhibited, rescued both nuclear elongation and CD69 expression. In addition, fibroblast cells plated on fibronectin micropatterns of different sizes showed correlation between nuclear shape index and tenascin C expression. Upon inhibiting the signaling intermediate Erk, tenascin C expression was down regulated although the nuclear shape index remained unaltered. Our results highlight the importance of specific signaling intermediates accompanied with nuclear deformation in the modulation of cellular genomic programs

Kinetic measurement of ribosome motor stalling force

We measure the ribosome motor stalling forces to unzip mRNA polymers during gene expression. An approach of using the changes in the reaction rate constants to determine the molecular motor forces is presented. Specific antisense DNA oligomers complementary to mRNA templates are used as kinetic barriers for estimating the ribosome forces using real time bioluminescence detection of luciferase gene expression. The rate constants are determined by comparing the experimental data with numerical simulation of gene expression to deduce the ribosome force (26.5±1 pN) required to unzip mRNA polymers. Understanding the forces generated by the ribosome may also enable the construction of information-based artificial nanoscale machines

Distinct levels in the nanoscale organization of DNA-histone complex revealed by its mechanical unfolding

Mechanical unfolding of nanoscale DNA-histone complex, using an atomic force microscope, shows a stepwise disassembly of histones from the nucleosome. A quantitative analysis of the rupture jump statistics and the length released per jump reveals insights into the possible histone contacts within the octamer complex. The measured ruptures correlate with the breakage of multiple contacts that stabilize the histone octamer. These results provide a mechanistic basis by which stepwise disassembly of histone proteins may result from an external force exerted by the adenosinetriphosphate (ATP) dependent chromatin remodeling machines to access regulatory sites on DNA