Changes In DNA Structure Research Articles

Autophagy receptor NDP52 alters DNA conformation to modulate RNA polymerase II transcription

NDP52 is an autophagy receptor involved in the recognition and degradation of invading pathogens and damaged organelles. Although NDP52 was first identified in the nucleus and is expressed throughout the cell, to date, there is no clear nuclear functions for NDP52. Here, we use a multidisciplinary approach to characterise the biochemical properties and nuclear roles of NDP52. We find that NDP52 clusters with RNA Polymerase II (RNAPII) at transcription initiation sites and that its overexpression promotes the formation of additional transcriptional clusters. We also show that depletion of NDP52 impacts overall gene expression levels in two model mammalian cells, and that transcription inhibition affects the spatial organisation and molecular dynamics of NDP52 in the nucleus. This directly links NDP52 to a role in RNAPII-dependent transcription. Furthermore, we also show that NDP52 binds specifically and with high affinity to double-stranded DNA (dsDNA) and that this interaction leads to changes in DNA structure in vitro. This, together with our proteomics data indicating enrichment for interactions with nucleosome remodelling proteins and DNA structure regulators, suggests a possible function for NDP52 in chromatin regulation. Overall, here we uncover nuclear roles for NDP52 in gene expression and DNA structure regulation.

DNA Morphology: Global Alteration of DNA Topological States Induced by Chemotherapeutic Agents and Its Implication in Cancer.

The dynamic topological states of chromosomal DNA regulate many cellular fundamental processes universally in all three domains of life, that is, bacteria, archaea, and eukaryotes. DNA-binding proteins maintain the regional and global supercoiling of the chromosome and thereby regulate the chromatin architecture that ultimately influences the gene expression network and other DNA-centric molecular events in various microenvironments and growth phases. DNA-binding small molecules are pivotal weapons for treating a wide range of cancers. Recent advances in single-molecule biophysical tools have uncovered the fact that many DNA-binding ligands not only alter the regional DNA supercoiling but also modulate the overall morphology of DNA. Here we provide insight into recent advances in atomic force microscopy (AFM) acquired DNA structural change induced by therapeutically important mono- and bis-intercalating anticancer agents as well as DNA-adduct-forming anticancer drugs. We also emphasize the growing evidence of the mechanistic relevance of changes in DNA topology in the anticancer cellular responses of DNA-targeting chemotherapeutic agents.

Characterizing the structural rearrangement of the bacterial nucleoid by Dps.

Bacteria are subject to a wide variety of environmental stresses which requires a robust stress response system to protect the cell. A key protein in Escherichia coli known for its numerous functions in stress response and cell maintenance is the DNA-binding protein from starved cells (Dps). The Dps protein protects DNA via two different mechanisms. First, Dps oxidizes iron to prevent the generation of hydroxyl radicals. Second, in response to stress and throughout the stationary phase of the growth cycle, Dps oligomers bind to DNA and multimerize further to create a Dps-DNA complex that compacts. To access how Dps affects this structural change of DNA to afford protection of the cell's genome, photoactivatable localization microscopy is employed to generate nanometer-scale maps of the nucleoid structure throughout the E. coli growth phases. These maps are attained by super-resolving the localizations of single molecules of the nucleoid-associated protein HUα, which non-specifically binds to DNA, tagged with the photoactivatable fluorescent protein PAmCherry. Super-resolution maps of the nucleoid structure are compared to maps of the nucleoid in E. coli strains with dps deleted. Tagging Dps and other major nucleoid associated proteins with PAmCherry provides the distribution of these proteins across the nucleoid to gain insight on how and if Dps interacts with other nucleoid associated proteins. By observing these structural changes over the growth period of E. coli, insight can be gained about how changes in DNA structure can lead to protection via Dps.

Handling DNA malfunctions by unsupervised machine learning model

The cell cycle is a rich field for research, especially, the DNA damage. DNA damage, which happened naturally or as a result of environmental influences causes change in the chemical structure of DNA. The extent of DNA damage has a significant impact on the fate of the cell in later stages.In this paper, we introduced an Unsupervised Machine learning Model for DNA Damage Diagnosis and Analysis. Mainly, we employed K-means clustering unsupervised machine learning algorithms. Unsupervised algorithms commonly draw conclusions from datasets by solely utilizing input vectors, disregarding any known or labeled outcomes. The model provided deep insight about DNA damage and exposes the protein levels for proteins when work together in sub-network model to deal with DNA damage occurrence, the unsupervised artificial model explained the sub-network biological model activities in regard to the changing in their concentrations in several clusters, they have been grouped in such as (0 - no damage, 1 - low, 2 - medium, 3 - high, and 4 - excess) DNA damage clusters.The results provided a rational and persuasive explanation for numerous important phenomena, including the oscillation of the protein p53, in a clear and understandable manner. Which is encouraging since it demonstrates that the K-means clustering approach can be easily applied to many similar biological systems, which aids in better understanding the key dynamics of these systems.

Influence of oxidative stress on drug-DNA binding: Microcalorimetric and mechanistic insights with anticancer drugs

Oxidative stress causing the chronic inflammation, mediates to deadly diseases such as cancer, diabetes, cardiovascular disease. Maintaining the ratio of reactive oxygen species generation to scavenging, thus the oxidative stress is important to have healthy cells. Oxidative stress not only causes the deadly diseases but also affect the conformation of biomolecules such as protein and DNA, even at lower concentrations. Binding of drugs to DNA is susceptible to these DNA structural or conformational changes. Here, we have reported the influence of oxidative stress on binding of anti-cancer drugsdoxorubicin hydrochloride (DOX),mitoxantrone dihydrochloride (MTX), capecitabine (CPT), and ethidium bromide (EtBr) with DNA. Quantitative insights of association of these anticancer drugs with DNA in presence of different environments, in vitro (pH – 7.4), cellular (100:1 ratio of GSH to GSSG) and oxidative stress condition (1:1 ratio of GSH to GSSG) have been compared. Microcalorimetry has been used for determination of thermodynamic parameters, binding constant Kb, molar enthalpy of interaction (ΔHm), molar entropy of interaction (ΔSm) & stoichiometry (n). Fluorescence spectroscopy has been used to illustrate the mode of drug-DNA binding and also to complement the ITC results. Insignificant changes in thermodynamic parameters for drug-DNA binding in in vitro and in presence of cellular condition were observed whereas significant decrease in binding affinity and stoichiometry was observed in the presence of oxidative stress conditions. Quantitative understanding of drug-DNA binding and influence of oxidative stress in terms of thermodynamic signature assists in the structure-based drug designing and development.

A Novel Biosensor for Detecting Vitamin C in Milk Powder Based on Hg2+- Mediated DNA Structural Changes

Background: Detection of Vitamin C (Vc) is very important to protect human health. A lot of methods have been developed for the detection of Vc. However, many methods require complex material preparation and skilled operators. Thus, a simple, label-free biosensor is still urgently needed. Methods: In this work, N-methylmesoporphyrin IX (NMM)/G-quadruplex pair was used as a labelfree signal reporter. Without Vc, the G-quadruplex DNA and its incomplete complementary chain could form a duplex structure by T-Hg(II)-T mismatch. In this case, the G-quadruplex structure could not be formed. When Vc was added, the Hg2+ was reduced to Hg(0). Then, the G-quadruplex DNA became free and formed a G-quadruplex structure to emit fluorescence signals. Results: Under optimal conditions, this biosensor showed a good linear response in the range of 0.2 - 4.0 μM and a low limit of detection (19.9 nM). This biosensor also had good selectivity towards Vc. Meanwhile, the satisfactory recovery rates (93.2%-102.8%) suggested that this biosensor had potential for measuring Vc in real samples. Conclusion: In this work, a simple label-free fluorescent biosensor for the detection of Vc based on Hg2+-mediated DNA structural changes had been developed. The whole experiment was simple and all reagents were commercialized. The label-free detection was realized by NMM/G-quadruplex as a signal reporter. This biosensor was very sensitive with a low limit of detection. It had a potential practical application for Vc detection in milk powder.

In vitro single molecule and bulk phase studies reveal the AP-1 transcription factor cFos binds to DNA without its partner cJun

The AP-1 transcription factor family crucially regulates progression of the cell cycle, as well as playing roles in proliferation, differentiation, and the stress response. The two best described AP-1 family members, cFos and cJun, are known to dimerize to form a functional AP-1 heterodimer that binds to a consensus response element sequence. Although cJun can also homodimerize and bind to DNA, the canonical view is that cFos cannot bind DNA without heterodimerizing with cJun. Here, we show that cFos can actually bind to DNA in the absence of cJun in vitro. Using dual color single molecule imaging of cFos alone, we directly visualize binding to and movement on DNA. Of all these DNA-bound proteins, detailed analysis suggested 30 to 46% were homodimers. Furthermore, we constructed fluorescent protein fusions of cFos and cJun for Förster resonance energy transfer experiments. These constructs indicated complete dimerization of cJun, but although cFos could dimerize, its extent was reduced. Finally, to provide orthogonal confirmation of cFos binding to DNA, we performed bulk-phase circular dichroism experiments that showed clear structural changes in DNA; these were found to be specific to the AP-1 consensus sequence. Taken together, our results clearly show cFos can interact with DNA both as monomers and dimers independently of its archetypal partner, cJun.

Size-dependent theoretical and experimental photothermal conversion efficiency of spherical gold nanoparticles

IntroductionDue to their biocompatible and plasmonic properties, gold nanoparticles (Au NPs) are good candidates to be photosensitizers in photothermal cancer therapy (PTT). Materials and methodsIn this paper, the dependence of the NIR-light-to-heat energy on Au NPs size was investigated. Moreover, to determine the photosensitizing properties of gold nanoparticles, PTT was conducted on two colon cell lines: SW480 and SW620 by irradiating them with two lasers having different wavelengths. ResultsTransmission electron microscopy showed that the respective sizes of Au NPs were 10 nm, 12 nm and 16 nm. Moreover, local as well as global structural measurements showed that all synthesized Au NPs were crystalline and UV-Vis spectroscopy revealed that with increasing nanoparticles size the position of the surface Plasmon resonance (SPR) peaks is shifted to higher wavelengths. Decrease of cells viability was observed, when they were cultured with Au NPs and irradiated by 650 nm and 808 nm lasers. Moreover, FTIR and Raman spectra of cells, showed structural changes in DNA, phospholipids, proteins and cholesterol caused by the addition of nanoparticles and laser irradiation. The chemical changes were more pronounced in the cells cultured with Au NPs and irradiated by 650 nm lasers and these changes were dependent on the nanoparticle size. Moreover, the viability of cells investigated by the MTS assay showed, that the percentage of dead cells (∼40%) is the highest for cells cultured with 8 nm Au NPs and irradiated by the 650 nm laser. The photothermal conversion efficiency calculated from the experimental results showed a decrease of this parameter from 70% to 55% and from 61% to 48% with increasing particle size, for 650 nm and 808 nm lasers, respectively. ConclusionsThe obtained results showed that the photothermal conversion efficiency of Au NPs is size-tunable, and can be correlated with the absorption/extinction ratios calculated by the Mie theory.

Extrachromosomal DNA in Cancer.

In cancer, complex genome rearrangements and other structural alterations, including the amplification of oncogenes on circular extrachromosomal DNA (ecDNA) elements, drive the formation and progression of tumors. ecDNA is a particularly challenging structural alteration. By untethering oncogenes from chromosomal constraints, it elevates oncogene copy number, drives intratumoral genetic heterogeneity, promotes rapid tumor evolution, and results in treatment resistance. The profound changes in DNA shape and nuclear architecture generated by ecDNA alter the transcriptional landscape of tumors by catalyzing new types of regulatory interactions that do not occur on chromosomes. The current suite of tools for interrogating cancer genomes is well suited for deciphering sequence but has limited ability to resolve the complex changes in DNA structure and dynamics that ecDNA generates. Here, we review the challenges of resolving ecDNA form and function and discuss the emerging tool kit for deciphering ecDNA architecture and spatial organization, including what has been learned to date about how this dramatic change in shape alters tumor development, progression, and drug resistance.

Alkyne-Tagged Raman Probes for Local Environmental Sensing by Hydrogen-Deuterium Exchange.

Alkyne-tagged Raman probes have shown high promise for noninvasive and sensitive visualization of small biomolecules to understand their functional roles in live cells. However, the potential for alkynes to sense cellular environments that goes beyond imaging remains to be further explored. Here, we report a general strategy for Raman imaging-based local environment sensing by hydrogen-deuterium exchange (HDX) of terminal alkynes (termed alkyne-HDX). We first demonstrate, in multiple Raman probes, that deuterations of the alkynyl hydrogens lead to remarkable shifts of alkyne Raman peaks for about 130 cm-1, providing resolvable signals suited for imaging-based analysis with high specificity. Both our analytical derivation and experimental characterizations subsequently establish that HDX kinetics are linearly proportional to both alkyne pKas and environmental pDs. After validating the quantitative nature of this strategy, we apply alkyne-HDX to sensing local chemical and cellular environments. We establish that alkyne-HDX exhibits high sensitivity to various DNA structures and demonstrates the capacity to detect DNA structural changes in situ from UV-induced damage. We further show that this strategy is also applicable to resolve subtle pD variations in live cells. Altogether, our work lays the foundation for utilizing alkyne-HDX strategy to quantitatively sense the local environments for a broad spectrum of applications in complex biological systems.

Investigation of a high-sensitive electrochemical DNA biosensor for determination of Idarubicin and studies of DNA-binding properties

Serious efforts have always been made to detect DNA molecules by fast, cost-effective, and susceptible instruments and clarify changes in DNA structure exposed to chemotherapy medicines. Accordingly, the present study endeavored to introduce a novel DNA biosensor fabricated by surface modification of pencil graphite electrodes using polypyrrole/La2O3 nanoparticles@snowflake-like Cu2S nanostructure composite, or PP/La2O3 NP@SF-L Cu2S NS composites, to detect ds-DNA molecules and Idarubicin (IDA). To prepare the proposed DNA biosensor, the ds-DNA was immobilized on the PP/La2O3 NP@SF-L Cu2S NS/PGE surface. IDA was detected electrochemically using differential pulse voltammetry (DPV). Subsequently, ultra-high sensitivity was reported for the biosensor of ds-DNA/PP/La2O3 NP@SF-L Cu2S NS/PGE towards the IDA so that the obtained limit of detection (LOD) was 1.3 nM with the linear range between 0.01 and 500.0 μM. The active reaction sites and admirable electrochemical activity associated with the nanocomposites were probably responsible for the excellent efficiency of the proposed biosensor, which accelerates electron transfer on the electrode surface and intensifies ds-DNA immobilization. The binding properties of IDA and DNA were investigated by molecular dynamic simulation (MDS), molecular docking (MD), and multi-spectroscopic analysis.

Topoisomerase: An Overview

The structure of DNA is a double-stranded helix, where the four bases are paired and stored in the center of this helix. The two strands of DNA are intertwined and this would require the two strands to be untwisted in order to access the information stored. Topoisomerases catalyze and guide the unknotting of DNA by creating transient breaks in the DNA using a conserved Tyrosine as the catalytic residue. Two classes of Topoisomerses are identified yet. Since the overall chemical composition and connectivity of the DNA does not change, the tangled and untangled DNAs are chemical isomers, differing only in their global topology, hence the enzymes are named as Topoisomerases. The insertion of viral DNA into chromosomes and other forms of recombination also require the action of topoisomerases. Topoisomerase inhibitors are agents designed to interfere with the action of topoisomerase enzymes, which control the changes in DNA structure by catalyzing the breaking and rejoining of the phosphodiester backbone of DNA strands during the normal cell cycle. Thus they are found to be important tools for treatment of cancer.

Usnic acid induced changes in biomolecules and their association with apoptosis in squamous carcinoma (A-431) cells: A flow cytometry, FTIR and DLS spectroscopic study

Many natural products induce apoptotic cell death in cancer cells, though studies on their interactions with macromolecules are limited. For the first time, this study demonstrated the cytotoxic potential of usnic acid (UA) against squamous carcinoma (A-431) cells and the associated changes in cell surface proteins, lipids and DNA by attenuated total reflection- fourier transform infrared spectroscopy (ATR-FTIR) and dynamic light scattering (DLS) spectroscopic studies. The IC50 for UA was 98.9 µM after treatment of A-431 cells for 48 h, while the IC50 reduced to 39.2 µM after 72 h of incubation time. UA induced oxidative stress in treated cells as confirmed by DCFHDA flow cytometry assay, depletion in reduced glutathione and increase in lipid peroxidation. The oxidative stress resulted in conformation change in amide I, amide II protein bands and DNA as observed by ATR-FTIR in UA treated A-431 cells. Shift in secondary structures of proteins from α helix to β sheets and structural changes in DNA was observed in UA treated A-431 cells. An increase in the band intensity of phospholipids, increased distribution of lipid and change in membrane potential was noted in UA treated cells, which was confirmed by externalization of phosphatidylserine to the outer membrane by annexin V-FITC/PI assay. Increase in mitochondrial membrane potential, cell cycle arrest at G0/G1 phase by flow cytometry and activation of caspase-3/7 dependent proteins confirmed the UA induced apoptosis in treated A-431 cells. FTIR and DLS spectroscopy confirmed the changes in biomolecules after UA treatment, which were associated with apoptosis, as observed by flow cytometry.

Evaluation of the binding effect and cytotoxicity assay of 2-Ethyl-5-(4-methylphenyl) pyramido pyrazole ophthalazine trione on calf thymus DNA: spectroscopic, calorimetric, and molecular dynamics approaches.

With advances in new drug therapies, it is essential to understand the interactions between drugs and target molecules. In this study, we applied multiple spectroscopic techniques including absorbance, fluorescence, circular dichroism spectroscopy, viscosity, thermal melting, calorimetric, and molecular dynamics (MD) simulation to study the interaction between 2-Ethyl-5-(4-methylphenyl) pyramido pyrazole ophthalazine trione (PPF) and calf thymus DNA (ct DNA) in the absence or presence of histone H1. PPF exhibits a high binding affinity towards ct DNA in binary and ternary systems. In addition, the result for the binding constant was observed within the range 104 M-1 achieved through fluorescence quenching data, while the values for enthalpy and entropy changes for ct DNA-PPF and (ct DNA-H1) PPF complexes were measured to be -72.54kJ.mol-1 , -161.14J.mol-1 K-1 , -85.34kJ.mol-1 , and -19.023J.mol-1 K-1 , respectively. Furthermore, in accordance with circular dichroism spectra, the inducement of ct DNA structural changes was observed during binding of PPF and H1 in binary and ternary system forms. The essential roles of hydrogen bonding and van der Waals forces throughout the interaction were suggested using thermodynamic parameters. According to the obtained data, the interaction mode of ct DNA-PPF and (ct DNA-H1) PPF complexes was intercalation binding. Suggested by the MD simulation study, the ct DNA-H1 complex caused a reduction in the stability of the DNA structure in the presence or absence of ligand, which demonstrated that PPF as an intercalating agent can further distort the structure. The information achieved from this study will be very helpful in understanding the effects of PPF on the conformational state of ct DNA in the absence or presence of the H1 molecule, which seems to be quite significant for clarifying the mechanisms of action and its pharmacokinetics.

Development of a novel liquid crystal Apta-sensing platform using P-shape molecular switch

Described herein is a liquid crystal (LC)-based aptasensor via employing the reorientation of LC triggered by the conformational changes of a P-shaped DNA structure. The structure consists of a short linker sequence as an immobilizer probe with ability to hybridize with the central part of the intact aptamer (Apt) sequence and an Apt terminal-locker (ATL) strand with complementary segments of the Apt terminal fragments. Bindings of two arm segments of the Apt sequence with the ATL strand enforces it to form a P-shaped configuration on the sensing platform. The selective interaction between the Apt strand and OTA leads to the disassembly of the Apt-ATL hybrid, collapse of the P-shaped structure, and consequently, transition of the optical appearance of the aptasensor texture. Determination of Ochratoxin A (OTA) in foods is an urgent demand in attempt to minimize food safety risks. To demonstrate the feasibility of our aptasensing design, the OTA specific aptamer was selected as a model. The developed LC aptasensor possesses a wide linear range from 0.01 aM to 100 pM, ultra-low limit of detection (LOD) of 0.0078 aM, and quantitative recoveries of 91–103.51% for OTA in rice and grape juice samples. This study proposes a novel and universal LC-based platform for facile, ultra-sensitive, and precision sensing of hazardous analytes in real samples.

Synergetic Inactivation Mechanism of Protocatechuic Acid and High Hydrostatic Pressure against Escherichia coli O157:H7.

With the wide application of high hydrostatic pressure (HHP) technology in the food industry, safety issues regarding food products, resulting in potential food safety hazards, have arisen. To address such problems, this study explored the synergetic bactericidal effects and mechanisms of protocatechuic acid (PCA) and HHP against Escherichia coli O157:H7. At greater than 200 MPa, PCA (1.25 mg/mL for 60 min) plus HHP treatments had significant synergetic bactericidal effects that positively correlated with pressure. After a combined treatment at 500 MPa for 5 min, an approximate 9.0 log CFU/mL colony decline occurred, whereas the individual HHP and PCA treatments caused 4.48 and 1.06 log CFU/mL colony decreases, respectively. Mechanistically, membrane integrity and morphology were damaged, and the permeability increased when E. coli O157: H7 was exposed to the synergetic stress of PCA plus HHP. Inside cells, the synergetic treatment additionally targeted the activities of enzymes such as superoxide dismutase, catalase and ATPase, which were inhibited significantly (p ≤ 0.05) when exposed to high pressure. Moreover, an analysis of circular dichroism spectra indicated that the synergetic treatment caused a change in DNA structure, which was expressed as the redshift of the characteristic absorption peak. Thus, the synergetic treatment of PCA plus HHP may be used as a decontamination method owing to the good bactericidal effects on multiple targets.

Kinetics of DNA looping by Anabaena sensory rhodopsin transducer (ASRT) by using DNA cyclization assay

DNA cyclization assay together with single-molecule FRET was employed to monitor protein-mediated bending of a short dsDNA (~ 100 bp). This method provides a simple and easy way to monitor the structural change of DNA in real-time without necessitating prior knowledge of the molecular structures for the optimal dye-labeling. This assay was applied to study how Anabaena sensory rhodopsin transducer (ASRT) facilitates loop formation of DNA as a possible mechanism for gene regulation. The ASRT-induced DNA looping was maximized at 50 mM of Na+, while Mg2+ also played an essential role in the loop formation.

Changes in gene-gene interactions associated with cancer onset and progression are largely independent of changes in gene expression

SummaryRecent findings indicate that changes underlying cancer onset and progression are not only attributable to changes in DNA structure and expression of individual genes but to changes in interactions among these genes as well. We examined co-expression changes in gene-network structure occurring during the onset and progression of nine different cancer types. Network complexity is generally reduced in the transition from normal precursor tissues to corresponding primary tumors. Cross-tissue cancer network similarity generally increases in early-stage cancers followed by a subsequent loss in cross-tissue cancer similarity as tumors reacquire cancer-specific network complexity. Gene-gene connections remaining stable through cancer development are enriched for “housekeeping” gene functions, whereas newly acquired interactions are associated with established cancer-promoting functions. Surprisingly, >90% of changes in gene-gene network interactions in cancers are not associated with changes in the expression of network genes relative to normal precursor tissues.

Large-scale DNA demethylation occurs in proliferating ovarian granulosa cells during mouse follicular development

During ovarian follicular development, granulosa cells proliferate and progressively differentiate to support oocyte maturation and ovulation. To determine the underlying links between proliferation and differentiation in granulosa cells, we determined changes in 1) the expression of genes regulating DNA methylation and 2) DNA methylation patterns, histone acetylation levels and genomic DNA structure. In response to equine chorionic gonadotropin (eCG), granulosa cell proliferation increased, DNA methyltransferase (DNMT1) significantly decreased and Tet methylcytosine dioxygenase 2 (TET2) significantly increased in S-phase granulosa cells. Comprehensive MeDIP-seq analyses documented that eCG treatment decreased methylation of promoter regions in approximately 40% of the genes in granulosa cells. The expression of specific demethylated genes was significantly increased in association with specific histone modifications and changes in DNA structure. These epigenetic processes were suppressed by a cell cycle inhibitor. Based on these results, we propose that the timing of sequential epigenetic events is essential for progressive, stepwise changes in granulosa cell differentiation.

Substituent effect of benzyl moiety in nitroquinoxaline small molecules upon DNA binding: Cumulative destacking of DNA nucleobases leading to histone eviction

Cooperative disruption of Watson-Crick hydrogen bonds, as well as base-destacking, is shown to be triggered by a quinoxaline-based small molecule consisting of an N,N-dimethylaminopropyl tether, and a para-substituted benzyl moiety. This events lead to superstructure formation and DNA condensation as evident from biophysical experiments and classical molecular dynamics simulations. The DNA superstructure formation by mono-quinoxaline derivatives is highly entropically favored and predominantly driven by hydrophobic interactions. Furthermore, oversupercoiling of DNA and base-destacking cumulatively induces histone eviction from in-vitro assembled nucleosomes at lower micromolar concentrations implicating biological relevance. The DNA structural modulation and histone eviction capacity of the benzyl para-substituents are in the order: –I > –CF3> -Br > -Me > -OMe > –OH, which is largely guided by the polarity of benzyl para-substituent and the resulting molecular topology. The most hydrophobic derivative 3c with para-iodo benzyl moiety causes maximal disruption of base pairing and generation of superstructures. Both these events gradually diminish as the polarity of the benzyl para-substituent increases. On the other hand, quinoxaline derivatives having heterocyclic ring instead of benzyl ring, or in the absence of N,N-dimethylamino head-group, is incapable of inducing any DNA structural change and histone eviction. Further, the quinoxaline compounds displayed potent anticancer activities against different cancer cell lines which directly correlates with the hydrophobic effects of the benzyl para-substituents. Overall, the present study provides new insights into the mechanistic approach of DNA structural modulation driven histone eviction guided by the hydrophobicity of synthesized compounds leading to cellular cytotoxicity towards cancer cells.