Biology subjects

Padinhateeri, R.

Publications and source records attributed to Padinhateeri, R..

25 records · Page 2Linked to original sources

bioRxiv2020

Continuous variable response, kinetic gating and connectivity that determine topology of insulin signaling are perturbed in hyper-insulinemic states

Understanding kinetic control of biological processes is as important as identifying components that constitute pathways. Insulin signaling (IS) is central for almost all metazoans and its perturbations are associated with various diseases and aging. While temporal phosphorylation changes and kinetic constants have provided some insights, constant or variable parameters that establish and maintain signal topology are poorly understood. Our iterative experimental and mathematical simulation-based approaches reveal novel kinetic parameters of IS that encode concentration and nutrient dependent information. Further, we find that pulsatile fasting insulin rewires IS akin to memory and in anticipation of a fed response. Importantly, selective kinetic gating of signals and maximum connectivity, between metabolic and growth-factor arms under normo-insulinemic states, maintains network topology. In addition to unraveling kinetic constraints that determine cascade architecture, our findings will help in identifying novel therapeutic strategies that conserve coupling between metabolic and growth-factor arms, which is lost in diseases and conditions of hyperinsulinemia.

systems biology↗

bioRxiv2019

The accidental ally: Nucleosomal barriers can accelerate cohesin mediated loop formation in chromatin

An important question in the context of the 3D organization of chromosomes is the mechanism of formation of large loops between distant base pairs. Recent experiments suggest that the formation of loops might be mediated by Loop Extrusion Factor proteins like cohesin. Experiments on cohesin have shown that cohesins walk diffusively on the DNA, and that nucleosomes act as obstacles to the diffusion, lowering the permeability and hence reducing the effective diffusion constant. An estimation of the times required to form the loops of typical sizes seen in Hi-C experiments using these low effective diffusion constants leads to times that are unphysically large. The puzzle then is the following, how does a cohesin molecule diffusing on the DNA backbone achieve speeds necessary to form the large loops seen in experiments? We propose a simple answer to this puzzle, and show that while at low densities, nucleosomes act as barriers to cohesin diffusion, beyond a certain concentration, they can reduce loop formation times due to a subtle interplay between the nucleosome size and the mean linker length. This effect is further enhanced on considering stochastic binding kinetics of nucleosomes on the DNA backbone, and leads to predictions of lower loop formation times than might be expected from a naive obstacle picture of nucleosomes.

biophysics↗

bioRxiv2019

Computing 3D chromatin configurations from contact probability maps by Inverse Brownian Dynamics

The three-dimensional organization of chromatin, on the length scale of a few genes, is crucial in determining the functional state -- accessibility and the amount of gene expression -- of the chromatin. Recent advances in chromosome conformation capture experiments provide partial information on the chromatin organization in a cell population, namely the contact count between any segment pairs. However, given the contact matrix, determining the complete 3D organization of the whole chromatin polymer is an inverse problem. In the present work, an Inverse Brownian Dynamics (IBD) method has been proposed to compute the optimal interaction strengths between different segments of chromatin such that the experimentally measured contact count probability constraints are satisfied. Applying this method to the -globin gene locus in two different cell types, we predict the 3D organizations corresponding to active and repressed states of chromatin at the locus. We show that the average distance between any two segments of the region has a broad distribution and cannot be computed as a simple inverse relation based on the contact probability alone. We also address the normalization problem of the contact count matrix and argue that extra measurements of polymer properties such as radius of gyration may be required to resolve the problem.\n\nSIGNIFICANCEChromosome conformation capture experiments such as 5C and Hi-C provide information on the contact counts between different segments of chromatin, but not the interaction strengths that lead to these counts. Here a methodology is proposed by which this inverse problem can be solved, namely, given the contact probabilities between all segment pairs, what is the pair-wise interaction strength that leads to this value? With the knowledge of pair-wise interactions determined in this manner, it is then possible to evaluate the 3D organization of chromatin and to determine the true relationship between contact probabilities and spatial distances.

biophysics↗

bioRxiv2019

Regulation of Microtubule Disassembly by Spatially Heterogeneous Patterns of Acetylation

Microtubules (MTs) are bio-polymers, composed of tubulin proteins, involved in several functions such as cell division, transport of cargoes within cells, maintaining cellular structures etc. Their kinetics are often affected by chemical modifications on the filament known as Post Translational Modifications (PTMs). Acetylation is a PTM which occurs on the luminal surface of the MT lattice and has been observed to reduce the lateral interaction between tubulins on adjacent protofilaments. Depending on the properties of the acetylase enzyme TAT1 and the structural features of MTs, the patterns of acetylation formed on MTs are observed to be quite diverse. In this study, we present a multi-protofilament model with spatially heterogenous patterns of acetylation, and investigate how the local kinetic differences arising from heterogeneity affect the global kinetics of MT filaments. From the computational study we conclude that a filament with spatially uniform acetylation is least stable against disassembly, while ones with more clustered acetylation patterns may provide better resistance against disassembly. The increase in disassembly times for clustered pattern as compared to uniform pattern can be upto fifty percent for identical amounts of acetylation. Given that acetylated MTs affect several cellular functions as well as diseases such as cancer, our study indicates that spatial patterns of acetylation need to be focussed on, apart from the overall amount of acetylation.\n\nAuthor SummaryMicrotubules (MTs) form a crucial part of the cytoskeletal machinery which regulates several cellular processes. The basic building block of MTs are tubulin proteins. These proteins assemble in lateral and longitudinal directions to form a hollow cylindrical structure of a MT. There are chemical modifications on tubulin, known as Post Translational Modifications (PTMs), which affect the stability and dynamics of MT filaments. We computationally study how one such PTM, namely acetylation, affects the kinetics of disassembly of a MT filament. We propose a model which incorporates spatially heterogeneous patterns of acetylation on MT filament and study how they may regulate the disassembly times and velocities, a factor hitherto unexplored in studies. We conclude that there are significant differences of disassembly velocities and their fluctuations depending on the differnces in spatial patterns of acetylation.

biophysics↗

bioRxiv2019

Liquid-liquid phase separation and liquid-to-solid transition mediate α-synuclein amyloid fibril containing hydrogel formation

-Synuclein (-Syn) aggregation and amyloid formation is directly linked with Parkinsons disease (PD) pathogenesis. However, the early events involved in this process remain unclear. Here, using in vitro reconstitution and cellular model, we show that liquid-liquid phase separation (LLPS) of -Syn precedes its aggregation. In particular, in vitro generated -Syn liquid-like droplets eventually undergo a liquid-to-solid transition and form amyloid-hydrogel containing oligomers and fibrillar species. Factors known to aggravate -Syn aggregation such as low pH, phosphomimic substitution, and familial PD mutation also promote -Syn LLPS and its subsequent maturation. We further demonstrate -Syn liquid droplet formation in cells, under oxidative stress. These cellular -Syn droplets eventually transform into perinuclear aggresomes, the process regulated by microtubules. The present work provides detailed insights into the phase separation behavior of natively unstructured -Syn and its conversion to a disease-associated aggregated state, which is highly relevant in PD pathogenesis.

neuroscience↗

bioRxiv2018

Irregular chromatin: packing density, fiber width and occurrence of heterogeneous clusters

How chromatin is folded in the lengthscale of a gene is an open question. Recent experiments have suggested that, in vivo, chromatin is folded in an irregular manner and not as an ordered fiber with a width of 30 nm expected from theories of higher order packaging. Using computational methods, we examine how the interplay between DNA-bending non histone proteins, histone tails, intra-chromatin electrostatic and other interactions decide the nature of packaging of chromatin. We show that while the DNA-bending non histone proteins make the chromatin irregular, they may not alter the packing density and size of the fiber. We find that the length of the interacting region and intra-chromatin electrostatic interactions influence the packing density, clustering of nucleosomes, and the width of the chromatin fiber. Our results suggest that the actively maintained heterogeneity in the interaction pattern will play an an important role in deciding the nature of packaging of chromatin.

biophysics↗

bioRxiv2017

Coupling of replisome movement with nucleosome dynamics can contribute to the parent-daughter information transfer

Positioning of nucleosomes along the genomic DNA is crucial for many cellular processes that include gene regulation and higher order packaging of chromatin. The question of how nucleosome-positioning information from a parent chromatin gets transferred to the daughter chromatin is highly intriguing. Accounting for experimentally known coupling between replisome movement and nucleosome dynamics, we propose a model that can explain the inheritance of nucleosome positioning. Simulating nucleosome dynamics during replication we argue that short pausing of the replication fork, associated with nucleosome disassembly, can be the event crucial for communicating nucleosome positioning information from parent to daughter. We show that the interplay of timescales between nucleosome disassembly ({tau}p) at the replication fork and nucleosome sliding behind the fork ({tau}s) can give rise to a rich \"phase diagram\" having different inherited patterns of nucleosome organization. Our model predicts that only when{tau} p [≥]{tau} s the daughter chromatin can inherit the precise nucleosome positioning of the parent.

biophysics↗