Biology subjects

Giri, R.

Publications and source records attributed to Giri, R..

At least 19 recordsLinked to original sources

bioRxiv2026

The microbiome determines the phenotype in CTLA-4 insufficient mice and men

CTLA-4 (haplo)insufficiency displays incomplete penetrance and phenotypic heterogeneity, indicating the involvement of additional disease modifiers beyond the genetic defect. Microbiome analyses reveal a positive association between disease severity and intestinal dysbiosis, highlighting the microbiome as a critical contributor. To investigate this relationship mechanistically, we generated Ctla4/- wildlings harboring a natural microbiota. Unlike specific pathogen free (SPF) counterparts, which remain healthy, Ctla4/- wildlings spontaneously develop disease phenotypes resembling human CTLA-4 haploinsufficiency. Disease onset is followed by reduced microbial diversity and expansion of pathobionts. Integrative immunophenotyping shows that the natural microbiota synergizes with Ctla4 haploinsufficiency to reshape innate and adaptive immune compartments, generating a sustained pro-inflammatory milieu and reduced CTLA-4 expression in the cecum. Furthermore, microbiota-derived metabolites promote inflammatory cytokine production in both murine and human primary T cells via NF-{kappa}B activation. Collectively, Ctla4/- wildlings constitute an effective model for dissecting microbiome-immune crosstalk in CTLA-4 (haplo)insufficiency and for exploring therapeutic strategies.

immunology↗

bioRxiv2026

Microbial Immunomodulatory Function is Altered in Inflammatory Bowel Disease Independent of Microbial Composition

Inflammatory bowel diseases (IBD) are chronic and relapsing immune-mediated conditions in which the NF-{kappa}B and STAT3 signalling pathways play central roles in pathogenesis. While gut microbiome dysbiosis is well described in IBD, its functional consequences, particularly microbial immunomodulatory function (MIF) remain poorly defined. Here, we integrated functional assays with metagenomic profiling to characterise microbiome-driven immune modulation in IBD. Faecal water from non-IBD controls and IBD patients was assessed using NF-{kappa}B and STAT3 reporter systems, alongside high-throughput screening of 2,820 bacterial isolates (94 per subject). Despite minimal differences in overall microbial composition, faecal water from patients with ulcerative colitis (UC) and Crohns disease (CD) significantly increased NF-{kappa}B activity under both basal and TNF-stimulated conditions. Non-IBD controls harboured a higher proportion of suppressive bacterial isolates (21.52%) compared to UC (5.64%) and CD (2.45%), whereas activating isolates were enriched in IBD (UC: 34.47%; CD: 39.36% vs. non-IBD: 5.35%). Similar trends were observed for IL-23-mediated STAT3 activation. Integration of metagenomic and pathway-level analyses identified 24 KEGG pathways associated with inflammatory signalling and faecal lipocalin-2, as well as amino acid, carbohydrate, lipid, cofactor, and nucleotide metabolism. These findings demonstrate that the IBD microbiome is functionally reprogrammed toward pro-inflammatory activity, independent of compositional changes. Defining patient-specific MIF profiles may provide a framework for precision microbiome-based therapeutic strategies in IBD.

microbiology↗

bioRxiv2026

Bioactive produced by Enterococcus Faecalis targets IL-23 signalling and protects against colitis and joint disease

The IL-23/Th17 axis is a central driver of intestinal and spondyloarthritic inflammation, yet upstream regulatory mechanisms linking microbial signals to IL-23 production remain incompletely defined. NF-{kappa}B signalling, particularly via the c-Rel subunit, is a critical transcriptional regulator of IL23A (p19), positioning c-Rel as a nodal checkpoint in mucosal inflammation. Here, we demonstrate that cell-free supernatant derived from Enterococcus faecalis AHG0090 (AHG0090-CS) suppresses c-Rel-dependent IL-23 signalling and attenuates inflammatory pathology across murine models of gut and joint disease. In the ZAP-70 mutant SKG model of spondyloarthritis and ileitis, AHG0090-CS significantly reduced weight loss, joint scores, and histological gut inflammation following curdlan challenge. In the Winnie model of spontaneous colitis, treatment similarly diminished inflammatory cytokine production. Mechanistically, AHG0090-CS reduced IL-23p19 mRNA and protein expression in intestinal tissue and lamina propria myeloid cells, accompanied by decreased nuclear c-Rel intensity. Suppression extended to downstream IL-23-associated cytokines including IL-17A, GM- CSF, MCP-1 and IL-6. In human peripheral blood mononuclear cells and macrophages, AHG0090-CS attenuated LPS-induced IL-23 and pro-inflammatory cytokine production, supporting translational relevance. Collectively, these findings identify microbial modulation of c-Rel-dependent IL-23 signalling as a tractable mechanism to restrain gut-joint inflammation and highlight targeting upstream NF-{kappa}B pathways as a therapeutic strategy in IL-23-driven immune-mediated disease.

microbiology↗

bioRxiv2026

Endotoxemia and TLR4 via tissue resident macrophages triggers anemia in mouse model of colitis

Anemia is one of the most debilitating and frequent complication of inflammatory bowel diseases (IBD) and is often treated with iron supplementation, which has limited efficacy and causes additional co-morbidities. Damaged intestinal barrier function is a hallmark of IBD and causes the translocation of endotoxins from gut bacteria into the bloodstream. A previous study in mice reported that endotoxins suppress erythropoiesis by reprogramming erythroblastic island macrophages (EBI M{varphi}). Here, we show that IBD patients and mice with acute colitis developed endotoxemia associated with anemia. Endotoxemia in IBD patients was negatively correlated with blood erythrocyte counts. In line with this, mice with acute colitis caused by drinking water containing dextrin sodium sulphate (DSS) had endotoxemia together with anemia characterized by reduced red blood cell counts, hemoglobin content and hematocrit, and reduced medullary erythropoiesis which was in part compensated by increased extramedullary erythropoiesis. As the endotoxin receptor TLR4 is expressed by CD169+ gut-resident macrophages as well as erythroid island macrophages in the bone marrow, we tested the hypothesis that TLR4 expressed by these CD169+ macrophages mediate both inflammatory colitis and anemia. Indeed, mice with a conditional deletion of the Tlr4 gene specifically in CD169+ tissue-resident macrophages were protected from DSS-induced anemia, inflammation and colitis. In addition, treatment with the TLR4 inhibitor C34 abated inflammation and anemia in DSS mice. These results suggest that endotoxins leaking from the inflamed gut may play a crucial role in IBD and associated anemia via CD169+ tissue-resident macrophages and that drugs targeting TLR4 may protect against IBD-associated anemia.

immunology↗

bioRxiv2025

PCR-Based Detection and Phylogenetic Analysis of Candidatus Liberibacter asiaticus in Citrus Orchards Across Nepal

Citrus greening disease, also known as huanglongbing (HLB), is caused by the gram-negative -proteobacteria Candidatus Liberibacter species. This disease poses a significant threat to citrus production worldwide, including in Nepal. This study aimed to conduct the diagnosis and phylogenetic analysis of the citrus greening pathogen in Nepal using both conventional PCR and computational methods. A total of 1,026 samples were collected from thirteen districts across six provinces in the country. PCR-based diagnosis was performed using the primer set Las606/LSS, which targets the 16S rRNA gene of Candidatus Liberibacter asiaticus. Additionally, 16S rRNA gene sequencing was performed using sanger sequencing for five samples collected from different geographical regions. The obtained sequences were deposited in GenBank, and a phylogenetic tree was constructed based on these sequences. Among the 1,026 samples tested, 255 were positive, indicating the widespread distribution of HLB across Nepal. All consensus sequences from Nepal showed strong evolutionary relatedness within the C. L. asiaticus cluster, displayed over 99% genetic similarity with reference sequences from various parts of the world. Phylogenetic analysis revealed that the Nepalese sequences were closely related to C. L. asiaticus sequences from India, such as OM522080.1 (Punjab) and MH473394.1 (Meerut). Additionally, sequences obtained from different regions of Nepal clustered closely together. The findings of this research provide valuable insights into the genetic diversity and prevalence of citrus greening disease, which will aid the development of better management strategies to combat this devastating disease. Furthermore, the sequence data deposited in GenBank will serve as a key reference for future molecular studies on citrus greening disease.

molecular biology↗

bioRxiv2024

A truncation mutant of adenomatous polyposis coli (APC) impairs apical cell extrusion through elevated epithelial tissue tension

Tissue tension encompasses the mechanical forces exerted on solid tissues within animal bodies, originating from various sources such as cellular contractility, interactions with neighbouring cells and the extracellular matrix. Emerging evidence indicates that an imbalance in tissue tension can influence structural organisation, homeostasis and potentially contribute to disease. For instance, heightened tissue tension can impede apical cell extrusion, leading to the retention of apoptotic or transformed cells. In this study, we investigate the potential role of adenomatous polyposis coli (APC) in modulating tissue tension. Our findings reveal that expression of an APC truncation mutant elevates epithelial tension via the RhoA/ROCK pathway. This elevation induces morphological alterations and hampers apoptotic cell extrusion in cultured epithelial cells and organoids, both of which could be mitigated by pharmacologically restoring the tissue tension. This raises the possibility that APC mutations may exert pathogenetic effects by altering tissue mechanics.

cell biology↗

bioRxiv2024

Curcumin Inhibits Zika Virus NS3 Helicase Activity

In the last decades, many efforts have been put forward in search of inhibitors/drugs specific to ZIKV-infected patients. These inhibitors include approved drugs, natural products, nucleosides, small molecules, and many others to inhibit Zika virus replication in cell and mouse models. In this regard, natural product curcumin has been widely explored in various disease conditions and found to have inhibition potential against ZIKV replication as well. Further, we investigated its inhibitory ability against the ZIKV NS3 helicase. We found that the curcumin interacts with the NS3 helicase domain by forming stable contacts at NTPase and RNA binding sites during molecular dynamic simulation. Furthermore, curcumin inhibits NTPase activity of NS3 helicase with an IC50 of 64.42 M. Altogether, the mechanistic insight into the action of curcumin will enlighten further drug discovery research in finding novel molecules.

biochemistry↗

bioRxiv2023

Signal Peptide of Human Serum Albumin (residues 1-18) Forms Amyloid-like Aggregates

Signal peptides (SPs) are essential tools in sorting or translocating the synthesized proteins. However, except few examples, their studies in isolation are quite limited. Here we asked a question of the aggregation propensity of signal peptide (residues 1-18) of Human Serum Albumin (HSA). Molecular dynamic simulations were used to acquire observations into the mechanism through which signal peptides of HSA self-aggregated in physiological conditions. Further, we confirmed our results by combining dye-based assays, atomic force microscopy, and transmission electron microscopy techniques. It was noted that the signal peptide of HSA (region 1-18) forms typical amyloid-like fibrils. Graphical O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=170 SRC="FIGDIR/small/563726v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@404ffaorg.highwire.dtl.DTLVardef@42e4caorg.highwire.dtl.DTLVardef@1ba942aorg.highwire.dtl.DTLVardef@556bee_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

bioRxiv2023

Japanese Encephalitis Virus: A pan-proteome analysis for aggregation propensities and in vitro validation with Capsid anchor and 2K peptide

Japanese encephalitis infection is a vector-borne disease caused by the flavivirus Japanese encephalitis virus (JEV). It is responsible of severe brain infection in humans worldwide. Given the ubiquitous nature of complications and tropism associated with Japanese encephalitis (JE) infection, a holistic understanding of its molecular mechanism is essential. The phenomenon of abnormal protein aggregation into pathogenic amyloids is now increasingly linked to multiple human diseases, also known as Amyloidosis. Most are neurodegenerative disorders but amyloidosis is not restricted to a specific organ or tissue type. The overlap of viral protein aggregation with human pathologies remains limited and it is gaining momentum, especially after the devastating Covid-19 pandemic. Therefore, in this study we have examined the likelihood of aggregation for the entire collection of proteins in JEV. Multiple independent web server tools were employed to scan for potential amyloid prone-regions (APRs), and it was followed by in vitro validation using two JEV transmembrane domains, Capsid anchor and 2K peptides. These synthetic viral peptides were introduced to artificial aggregation-inducing conditions and then analyzed using a different dye-based assays and microscopy methods confirming amyloid-like fibril structure formation. We found these aggregates cytotoxic to human neuronal cell line and membrane damaging to human blood derived RBCs. The aggregation kinetics of both peptides is enhanced in the presence of artificial membrane models and seeds of self and diabetes hallmark protein Amylin. Our findings thereby strongly suggest the possibility of JEV protein aggregation playing a vital role in its pathogenesis, opening up a broad scope of future study. Also, the interplay between JEV protein aggregation and initiation/progression of other proteopathies is possible and needs further exploration. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/556571v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@2956d0org.highwire.dtl.DTLVardef@28128org.highwire.dtl.DTLVardef@6d7faaorg.highwire.dtl.DTLVardef@d89f40_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2023

Coronaviruses Spike glycoprotein endodomains: the sequence and structure-based comprehensive study

The flexibility of proteins makes them available to interact with many biomolecules in the cell. Specifically, such interactions in viruses help them to perform more functions despite having a smaller genome. Therefore, these flexible regions can be exciting and essential targets to be explored for their role in pathogenicity and therapeutic developments as they achieve essential interactions. In the continuation with our previous study on disordered analysis of SARS-CoV-2 spike cytoplasmic tail (CTR), or endodomain, here we have explored the disordered potential endodomains of six other coronaviruses using multiple bioinformatics approaches and molecular dynamics simulations. Based on the comprehensive analysis of its sequence and structural composition, we report the varying disorder propensity in endodomains of spike proteins of coronaviruses. The observations of this study may help to understand the importance of spike glycoprotein endodomain and creating therapeutic interventions against them.

bioinformatics↗

bioRxiv2023

Met58 and di-acidic motif located at C-terminal region of SARS-CoV-2 ORF6 plays a crucial role in its structural conformations

Despite being mostly neglected in structural biology, the C-terminal Regions (CTRs) are studied to be multifunctional in humans as well as in viruses. Their role in cellular processes such as trafficking, protein-protein interactions, and protein-lipid interactions are known due to their structural properties. In our previous findings on SARS-CoV-2 Spike and NSP1 proteins, the C-terminal regions (CTRs) are observed to be disordered and experimental evidence showed a gain of structure properties in different physiological environments. In this line, we have investigated the structural dynamics of CTR (residues 38-61) of SARS-CoV-2 ORF6 protein, disrupting bidirectional transport between the nucleus and cytoplasm. Like Spike and NSP1-CTR, the ORF6-CTR is also disordered in nature but possesses gain of structure properties in minimal physiological conditions. As per studies, the residue such as Methionine at 58th position in ORF6 is critical for interaction with Rae1-Nup98. Therefore, along with M58, we have identified a few other mutations from the literature and performed extensive structure modelling and dynamics studies using computational simulations. The exciting revelations in CTR models provide evidence of its structural flexibility and possible capabilities to perform multifunctionality inside the host.

biophysics↗

bioRxiv2022

Discovery And Characterization of Small Molecule Inhibitors of Zika Virus Replication

Zika virus (ZIKV) is a flavivirus, and ZIKV infections in the past 15 years have been linked to Guillain-Barre syndrome and severe complications during pregnancy associated with congenital Zika syndrome. There are no approved therapies or vaccines for ZIKV. In recent years, advances in structure-based drug design methodologies have accelerated drug development pipelines for identifying promising inhibitory compounds against viral diseases. Among ZIKV proteins, NS2B-NS3 protease is an attractive target for antiviral drug development due to its vital role in the proteolytic processing of the single polyprotein. To find potential inhibitors against ZIKV, we used molecular docking at the NS2B-NS3 protease active site as a virtual screening approach with small molecules diverse scaffold-based library with rigorous druglikeness filters. The top-hit compounds with stable molecular dynamics trajectories were then subjected to in-vitro efficacy testing against ZIKV. In docking and molecular dynamics simulation studies, compound F1289-0194 showed stable binding to the NS2B-NS3 protease active site. Furthermore, viral load assays, immunofluorescence, and plaque reduction assays demonstrated that compound F1289-0194 significantly reduced ZIKV load and replication in Vero cells while maintaining cellular integrity. Thus, the compound F1289-0194 merits further investigation as a novel inhibitor against ZIKV replication.

cell biology↗

bioRxiv2022

Investigating the folding dynamics of NS2B protein of Zika virus

NS2B protein of the Zika virus acts as a co-factor for NS3 protease where only the cytosolic domain of NS2B is sufficient for the protease activity. At the same time, NS2B also involves in remodeling the NS3 protease structure. In isolation, we previously proved the NS2B cytosolic domain (residues 49-95) as a disordered type peptide conformation. Further, this study investigated the overall dynamics of NS2B full-length protein. Our Alphafold2 structure modeling system revealed surprising similarities between selected flavivirus NS2B proteins. This similarity reflects that the NS2B protein across flavivirus is conserved fold-wise. The MD simulation of Zika virus NS2B full-length protein shows that the cytosolic domain as a part of full-length protein is a disorder region supporting our previous experimental finding, which suggests the disordered nature of the cytosolic domain in isolation. Since the cytosolic domain of NS2B is essential for protease activity, we have also investigated the folding and dynamics of the NS2B cytosolic domain (residues 49-95) that shows the disorder to alpha helix transition in TFE. On the other hand, in the presence of SDS, macromolecular crowder like ficoll and PEG do not induce secondary structural change. This dynamics study could have implications for some unknown folds of the NS2B protein.

biophysics↗

bioRxiv2022

Structural dynamics of Zika Virus NS1 via a reductionist approach reveal the disordered nature of its beta roll domain in isolation

Flavivirus Non-structural 1 (NS1) protein performs multiple functions such as host immune evasion, interaction with complement system factors, membrane rearrangement, etc. Therefore, it is highly plausible that significant structural and folding dynamics of NS1 might play a role in its multifunctionality. The dimeric structures of NS1 of multiple flaviviruses, including Zika virus (ZIKV), are available. However, its domain-wise dynamics perspective has not been explored so far. Therefore, it is of utmost importance to understand the structural conformations of NS1 and its domains in isolation, possibly highlighting the implications on the overall NS1 protein dynamics. Here, we have employed extensively long molecular dynamic (MD) simulations to understand the role of monomer, dimer, and a reductionist approach in understanding the dynamics of the three structural domains (i.e., {beta}- roll, wing, and {beta}-ladder) in isolation. Further, we experimentally validated our findings using CD spectroscopy and confirmed the intrinsically disordered behavior of NS1 {beta}-roll in isolation and lipid mimetic environments. We also found that the {beta}-ladder domain is highly flexible during long simulations. Therefore, we believe this study may have implications for significant dynamics played by NS1 protein, specifically during oligomerization of NS1. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=154 SRC="FIGDIR/small/488568v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@1b2928org.highwire.dtl.DTLVardef@12b0b68org.highwire.dtl.DTLVardef@f9da77org.highwire.dtl.DTLVardef@175fbc1_HPS_FORMAT_FIGEXP M_FIG C_FIG Schematic representation of the ZIKV NS1 protein and the models that we have used in this study.

biophysics↗

bioRxiv2022

The aggregation potential of Zika virus proteome

The ability of human encoded soluble proteins to convert into amyloid fibrils is now recognized as a generic phenomenon in several human illnesses. Typically, such disease causal proteins/peptides consist of aggregation-prone regions (APR) that make them susceptible to misfolding and assemble into highly ordered {beta}-sheet rich fibrils, distinct from their native soluble state. Here, we show that the zika virus (ZIKV) consists of several such aggregation prone hotspots spread across its entire proteome. Using a combination of high-accuracy prediction tools, we identified APRs in both structural and non-structural proteins of ZIKV. Furthermore, we have experimentally validated the bioinformatic results by subjecting the ZIKV proteins and peptides to artificial aggregation inducing environment. Using a combination of dye-based assays (ThT and ANS) and microscopy techniques (HR-TEM and AFM), we further characterized the morphological features of amyloid-like fibrils. We found that Envelope domain III (EDIII) protein, NS1 {beta}-roll peptide, membrane-embedded signal peptide 2K, and cytosolic region of NS4B protein to be highly aggregating in the experimental setup. Our findings also pave the way for an extensive and detailed functional analysis of these predicted APRs in the future to enhance our understanding of the role played by amyloids in the pathogenesis of flavivirus. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=122 SRC="FIGDIR/small/485915v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@1bedf4dorg.highwire.dtl.DTLVardef@364676org.highwire.dtl.DTLVardef@14bf54aorg.highwire.dtl.DTLVardef@1d4fc19_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

bioRxiv2021

Single-cell protein analysis reveals metastable states during the transition to a sensory organ fate

Cell fate decisions can be envisioned as bifurcating dynamical systems, and the decision that Drosophila cells make during sensory organ differentiation has been described as such. We extended these studies by focusing on the Senseless protein, which orchestrates sensory cell fate transitions. Wing cells contain intermediate Senseless numbers prior to their fate transition, after which they express much greater numbers of Senseless molecules as they differentiate. However, the dynamics are inconsistent with it being a simple bistable system. Cells with intermediate Senseless are best modeled as residing in four discrete states, each with a distinct protein number and occupying a specific region of the tissue. Although the states are stable over time, the number of molecules in each state vary with time. The fold-change in molecule number between adjacent states is invariant and robust to absolute protein number variation. Thus, cells transitioning to sensory fates exhibit metastability with relativistic properties.

developmental biology↗

bioRxiv2021

Investigating the conformational dynamics of Zika virus NS4B protein

Zika virus (ZIKV) NS4B protein is a membranotropic protein having multifunctional roles such as evasion of host-immune system, and induction of host membrane rearrangement for viral genome replication and processing of polyprotein. Despite its versatile functioning, its topology and dynamics are not entirely understood. Presently, there is no NMR or X-ray crystallography-based structure available for any flaviviral NS4B protein. Therefore, in this study, we have investigated the structural dynamics of Zika Virus NS4B protein through 3D structure models using molecular dynamics (MD) simulations approach and experiments. Subsequently, we employed a reductionist approach to understand the dynamics of ZIKV NS4B protein. For this, we studied its N-terminal (residues 1-38), C-terminal (residues 194-251), and cytosolic (residues 131-169) regions in isolation. Further, we have performed experiments to prove the maximum dynamics in its cytosolic region. Using a combination of computational tools and circular dichroism (CD) spectroscopy, we validate the cytosolic region as an intrinsically disordered protein region (IDR). The microsecond-long all atoms molecular dynamics and replica-exchange MD simulations complement the experimental observations. We have also analysed its behaviour under the influence of differently charged liposomes and macromolecular crowding agents which may have significance on its overall dynamics. Lastly, we have proposed a ZIKV NS4B protein model illustrating its putative topology consisting of various membrane-spanning and non-membranous regions. HighlightsO_LIMicrosecond simulations of NS4B N-terminus and cytosolic regions exposed their dynamic nature. C_LIO_LIC-terminal region remains intact in presence lipid bilayer during 1 s simulations. C_LIO_LISpectroscopic results also reveal the cytosolic region as an intrinsically disordered region. C_LIO_LICytosolic region exhibits gain-of-structure property under helix inducing solvents. C_LI

biophysics↗

bioRxiv2021

Mitoxantrone dihydrochloride, an FDA approved drug, binds with SARS-CoV-2 NSP1 C-terminal

One of the major virulence factors of SARS-CoV-2, NSP1, is a vital drug target due to its role in host immune evasion through multiple pathways. NSP1 protein is associated with inhibiting host mRNA translation by binding to the small subunit of ribosome through its C-terminal region. Previously, we have shown the structural dynamics of NSP1 C-terminal region (NSP1-CTR) in different physiological environments. So, it would be very interesting to investigate the druggable compounds that could bind with NSP1-CTR. Here, in this article, we have performed the different spectroscopic technique-based binding assays of an anticancer drug Mitoxantrone dihydrochloride (MTX) against the NSP1-CTR. We have also performed molecular docking followed by computational simulations with two different forcefields up to one microsecond. Overall, our results have suggested good binding between NSP1-CTR and MTX and may have implications in developing therapeutic strategies targeting NSP1 protein of SARS-CoV-2.

biophysics↗