Biology subjects

Rajput, R.

Publications and source records attributed to Rajput, R..

7 recordsLinked to original sources

bioRxiv2025

Divergent myelopoiesis and macrophage polarization underlie host susceptibility to chronic Chlamydia trachomatis infection

Chronic genital infection with Chlamydia trachomatis is associated with immune suppression and pathogen persistence, but the mechanisms underlying this phenomenon remain poorly defined. Here, we show that chronic C. trachomatis genital infection differentially reprograms myelopoiesis and macrophage polarization in genetically distinct mouse strains. In BALB/c mice, infection expanded bone marrow progenitors with enhanced colony-forming potential but skewed their differentiation toward immunosuppressive M2 macrophages and myeloid-derived suppressor cells (MDSCs). These cells accumulated in the spleen, blood, and genital tract, secreted IL-10 and TGF{beta}, and suppressed T cell proliferation, thereby impairing IFN{gamma}-driven clearance. In contrast, C57BL/6 mice exhibited C/EBP{beta}-driven myelopoiesis favoring M1 polarization, elevated IL-12 and nitric oxide production, and robust Th1 responses that facilitated bacterial elimination. Together, these findings demonstrate that host genetic background dictates divergent innate programming during C. trachomatis infection, and that the balance between M1- and M2/MDSC-dominated myelopoiesis determines whether infection resolves or progresses to chronicity.

immunology↗

bioRxiv2024

Ribozyme-based biosensor for imaging gene expression in plants

Detection of gene expression in plants is critical for understanding the molecular basis of complex plant biosystems and plant responses to environmental stresses. Here, we report the development of a split ribozyme-based biosensor that enables in vivo visualization of gene expression in plants. We demonstrated the utility of this biosensor in transient expression experiments (i.e., leaf infiltration in Nicotiana benthamiana) to detect RNAs derived from transgenes and tobacco rattle virus, respectively. Furthermore, we successfully engineered a split ribozyme-based biosensor in Arabidopsis thaliana for in vivo visualization of endogenous gene expression at the cellular level. In addition, we developed a platform for easy incorporation of different reporters into the RNA biosensor.

synthetic biology↗

bioRxiv2024

Type-B response regulators MaRR_B9 and MaRR_B12 coordinate cytokinin-mediated negative regulation of anthocyanin biosynthesis in banana fruits

O_LIAnthocyanins are pigments responsible for vibrant plant colors and play vital roles in plant physiology. Comparing two banana varieties, grand naine (GN) and red banana (RB), which display significant contrast in anthocyanin pigmentation. C_LIO_LITranscriptomic analysis of GN and RB peel and pulp tissue highlighted cytokinin-specific response regulators, MaRR_B9 and MaRR_B12, as regulators of anthocyanin biosynthesis. C_LIO_LIThrough a combination of physiological, molecular, and biochemical analyses, we demonstrate that MaRR_B9 and MaRR_B12 exert direct regulatory control over key structural genes of anthocyanin biosynthesis, dihydroflavanol reductase (MaDFR) and anthocyanidin synthase (MaANS). MaRR_B9 and MaRRB_12 interact with the promoters of MaDFR and MaANS, repressing the concerned genes in vivo. Overexpression of MaRR_B9 and MaRR_B12 in banana fruits leads to a reduction in anthocyanin content, particularly that of the cyanidin derivative, along with altered expression patterns of MaDFR and MaANS. C_LIO_LIThus, the present study identifies novel regulators of anthocyanin biosynthesis in banana and provides further evidence that the cytokinin regulatory network modifies anthocyanin accumulation in plants. In conclusion, our findings reveal new molecular targets, in the form of MaRRs for the genetic manipulation of anthocyanin content and thereby for the development of stress-resilient and nutritionally enriched crop plants. C_LI

plant biology↗

bioRxiv2023

The HSF-DREB-MYB regulatory module regulates flavonol biosynthesis and flavonoid B-ring hydroxylation in banana

Plant flavonols act primarily as ultraviolet radiation absorbers, reactive oxygen species scavengers, and phytoalexins, and they contribute to biotic and abiotic stress tolerance in plants. Banana (Musa acuminata), an herbaceous monocot and important fruit crop, accumulates flavonol derivatives in different organs, including the edible fruit pulp. Although flavonol content varies greatly in different organs, the molecular mechanisms involving transcriptional regulation of flavonol synthesis in banana are not known. Here, we characterized three SG7-R2R3 MYB transcription factors (MaMYBFA1, MaMYBFA2, and MaMYBFA3) and their upstream regulators, heat shock transcription factor (MaHSF11) and dehydration responsive element binding factor (MaDREB1), to elucidate the molecular mechanism involved in transcriptional regulation of flavonol biosynthesis in banana. MaMYBFA positively regulate flavonol synthase 2 (MaFLS2) and downregulates MaFLS1. We show these transcription factors to be weak regulators of flavonol synthesis. Overexpression of MaHSF11 enhances flavonol contents, particularly that of myricetin, and promotes flavonol B-ring hydroxylation, which contributes to the diversity of flavonol derivatives. MaHSF11 directly interacts with the MaFLS1 and flavonoid 3', 5'-hydroxylase1 (MaF3'5'H1) promoters, both in vitro and in vivo. MaHSF11 activates the expression of MaDREB1 directly, which in turn regulates the expression of MaMYBFA3. Overall, our study elucidates a novel regulatory mechanism for flavonol synthesis in banana and suggests possible targets for genetic optimization to enhance nutritional value and stress responses in this globally important fruit crop. Significance statementOur study reveals that the three stress-responsive SG7 MYB factors, MaMYBFA1-3, are modest regulators of flavonol biosynthesis in banana. In contrast, the drought- and heat-responsive MaHSF11 plays a strong, positive role in this pathway, enhancing flavonol biosynthesis and flavonoid B-ring hydroxylation by upregulating key biosynthetic genes, such as MaFLS1 and MaF35H1, as well as MaDREB1, another regulator of flavonol biosynthesis in banana.

plant biology↗

bioRxiv2022

Flavonols contrary affect the interconnected glucosinolate and camalexin biosynthesis pathway in Arabidopsis thaliana

Flavonols are structurally and functionally diverse molecules playing roles in plant biotic and abiotic stress tolerance, auxin transport inhibition, pollen development, etc. Despite their ubiquitous occurrence in land plants and multifunctionality, the effect of perturbation of flavonol biosynthesis over global gene expression and pathways other than flavonoid biosynthesis has not been studied in detail. To understand the signaling role of different flavonol metabolites, herein, we used the flavonol deficient Arabidopsis thaliana loss-of-function mutant flavonol synthase1 (fls1-3) as object of study. Comparative transcriptome and metabolic profiling were used to study the effects of genetic flavonol deficiency and exogenous supplementation with flavonol derivatives (kaempferol, quercetin and rutin) on different cellular processes in the seedling. Various flavonol biosynthesis-related regulatory and structural genes were found to be up-regulated in the fls1-3 mutant which could be reversed by exogenous flavonol feeding. Our manifold comparative studies indicated the modulation of various biological processes and metabolic pathways by flavonols. Camalexin biosynthesis was found to be negatively regulated by flavonols. Interestingly, flavonols appeared to promote the accumulation of aliphatic glucosinolate through transcription factor-mediated up-regulation of biosynthesis genes. Overall, this study provides new insights into molecular mechanisms by which flavonols interfere with the relevant signal chains and their molecular targets and adds new knowledge to the expanding plethora of biological activity of flavonols in plants. SignificanceComparative transcriptome and metabolomic profiling of genetic flavonol deficiency and exogenous flavonol supplementation in A. thaliana seedlings, for the first-time revealed the inverse regulation of interconnected specialized metabolite pathways by flavonol aglycones, and -glycosides. Flavonols negatively regulate camalexin biosynthesis, while promoting the accumulation of aliphatic glucosinolates. Our study adds new insights into the expanding plethora of biological activity of flavonols in plants and will help to uncover the molecular mechanisms by which flavonols interfere with the relevant signal chains and their molecular targets.

plant biology↗

bioRxiv2022

Interplay between R2R3 MYB-type activators and repressors regulates proanthocyanidin biosynthesis in banana (Musa acuminata)

O_LIProanthocyanidins are oligomeric flavonoid pigments that promote plant disease resistance and benefit human health. However, the transcriptional regulatory network that fine-tunes proanthocyanidin biosynthesis in banana (Musa acuminata) fruit remains poorly understood. C_LIO_LIWe characterized two proanthocyanidin-specific R2R3 MYB activators (MaMYBPA1-MaMYBPA2) and four repressors (MaMYBPR1-MaMYBPR4) to elucidate the mechanisms underlying the transcriptional regulation of proanthocyanidin biosynthesis in banana. C_LIO_LIHeterologous expression of MaMYBPA1 and MaMYBPA2 partially complemented the Arabidopsis thaliana proanthocyanidin-deficient transparent testa2 mutant. MaMYBPA1 and MaMYBPA2 interacted physically with MaMYCs and bound the promoters of the genes encoding anthocyanin synthase, leucoanthocyanidin reductase, and anthocyanidin reductase in vitro and form functional MBW complexes with MaTTG1 in A. thaliana protoplasts to transactivated these promoters in vivo. C_LIO_LIOverexpression of MaMYBPAs alone or with MaMYC in banana fruits induced proanthocyanidin accumulation and the transcription of proanthocyanidin biosynthesis- related genes. MaMYBPR repressors are also shown to interact with MaMYCs, forming repressing MBW complexes, and diminished proanthocyanidin accumulation. Interestingly the overexpression of MaMYBPA induces the expression of MaMYBPR, indicating an agile regulation of proanthocyanidin biosynthesis via the formation of competitive MBW complex. Taken together, our results reveal regulatory modules of R2R3 MYB- that fine-tune proanthocyanidin biosynthesis and offer possible targets for genetic manipulation in banana. C_LI

plant biology↗

bioRxiv2020

Experimental glaucoma retinal proteomics identifies mutually exclusive and overlapping molecular characteristics with human glaucoma

Current evidence suggests that exposure to chronically induced intraocular pressure (IOP) leads to neurodegenerative changes in the inner retina. This study aimed to determine retinal proteomic alterations in a rat model of glaucoma and compared findings with human retinal proteomics changes in glaucoma reported previously. We developed an experimental glaucoma rat model by subjecting the rats to increased IOP (9.3{+/-}0.1 vs 20.8{+/-}1.6 mm Hg) by weekly microbead injections into the eye (8 weeks). The retinal tissues were harvested from control and glaucomatous eyes and protein expression changes analysed using multiplexed quantitative proteomics approach. Immunofluorescence was performed for selected protein markers for data validation. Our study identified 4304 proteins in the rat retinas. Out of these, 139 proteins were downregulated ([≤]0.83) while expression of 109 proteins was upregulated ([≥]1.2-fold change) under glaucoma conditions (p[≤]0.05). Computational analysis revealed reduced expression of proteins associated with glutathione metabolism, mitochondrial dysfunction/oxidative phosphorylation, cytoskeleton and actin filament organisation, along with increased expression coagulation cascade, apoptosis, oxidative stress and RNA processing markers. Further functional network analysis highlighted the differential modulation of nuclear receptor signalling, cellular survival, protein synthesis, transport and cellular assembly pathways. Alterations in crystallin family, glutathione metabolism and mitochondrial dysfunction associated proteins shared similarities between the animal model of glaucoma and the human disease condition. In contrast, the activation of the classical complement pathway and upregulation of cholesterol transport proteins, were exclusive to the human glaucoma. These findings provide insights into the neurodegenerative mechanisms that are specifically affected in the retina in response to chronically elevated IOP.

neuroscience↗