Tomato glycosyltransferase Twi1 plays a role in flavonoid glycosylation and defence against virus

Abstract Background: Secondary metabolites play an important role in the plant defensive response. They are produced as a defence mechanism against biotic stress by providing plants with antimicrobial and antioxidant weapons. In higher plants, the majority of secondary metabolites accumulate as glycoconjugates. Glycosylation is one of the commonest modifications of secondary metabolites, and is carried out by enzymes called glycosyltransferases. Results: Here we provide evidence that the previously described tomato wound and pathogen-induced glycosyltransferase Twi1 displays in vitro activity toward the coumarins scopoletin, umbelliferone and esculetin, and the flavonoids quercetin and kaempferol, by uncovering a new role of this gene in plant glycosylation. To test its activity in vivo, Twi1-silenced transgenic tomato plants were generated and infected with Tomato spotted wilt virus. The Twi1-silenced plants showed a differential accumulation of Twi1 substrates and enhanced susceptibility to the virus. Conclusions: Biochemical in vitro assays and transgenic plants generation proved to be useful strategies to assign a role of tomato Twi1 in the plant defence response. Twi1 glycosyltransferase showed to regulate quercetin and kaempferol levels in tomato plants, affecting plant resistance to viral infection.

Tomato glycosyltransferase Twi1 plays a role in flavonoid glycosylation and defence against virus

10.21203/rs.2.10162/v4 ◽

2019 ◽

Author(s):

Laura Campos ◽

María Pilar López-Gresa ◽

Diana Fuertes ◽

José María Bellés ◽

Ismael Rodrigo ◽

...

Keyword(s):

Secondary Metabolites ◽

Higher Plants ◽

Plant Defence ◽

In Vitro Assays ◽

Tomato Plants ◽

Abstract Background: Secondary metabolites play an important role in the plant defensive response. They are produced as a defence mechanism against biotic stress by providing plants with antimicrobial and antioxidant weapons. In higher plants, the majority of secondary metabolites accumulate as glycoconjugates. Glycosylation is one of the commonest modifications of secondary metabolites, and is carried out by enzymes called glycosyltransferases. Results: Here we provide evidence that the previously described tomato wound and pathogen-induced glycosyltransferase Twi1 displays in vitro activity toward the coumarins scopoletin, umbelliferone and esculetin, and the flavonoids quercetin and kaempferol, by uncovering a new role of this gene in plant glycosylation. To test its activity in vivo, Twi1-silenced transgenic tomato plants were generated and infected with Tomato spotted wilt virus. The Twi1-silenced plants showed a differential accumulation of Twi1 substrates and enhanced susceptibility to the virus. Conclusions: Biochemical in vitro assays and transgenic plants generation proved to be useful strategies to assign a role of tomato Twi1 in the plant defence response. Twi1 glycosyltransferase showed to regulate quercetin and kaempferol levels in tomato plants, affecting plant resistance to viral infection.

Tomato glycosyltransferase Twi1 plays a role in flavonoid glycosylation and defence against virus

BMC Plant Biology ◽

10.1186/s12870-019-2063-9 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 2

Author(s):

Laura Campos ◽

María Pilar López-Gresa ◽

Diana Fuertes ◽

José María Bellés ◽

Ismael Rodrigo ◽

...

Keyword(s):

Secondary Metabolites ◽

Higher Plants ◽

Plant Defence ◽

In Vitro Assays ◽

Tomato Plants ◽

Abstract Background Secondary metabolites play an important role in the plant defensive response. They are produced as a defence mechanism against biotic stress by providing plants with antimicrobial and antioxidant weapons. In higher plants, the majority of secondary metabolites accumulate as glycoconjugates. Glycosylation is one of the commonest modifications of secondary metabolites, and is carried out by enzymes called glycosyltransferases. Results Here we provide evidence that the previously described tomato wound and pathogen-induced glycosyltransferase Twi1 displays in vitro activity toward the coumarins scopoletin, umbelliferone and esculetin, and the flavonoids quercetin and kaempferol, by uncovering a new role of this gene in plant glycosylation. To test its activity in vivo, Twi1-silenced transgenic tomato plants were generated and infected with Tomato spotted wilt virus. The Twi1-silenced plants showed a differential accumulation of Twi1 substrates and enhanced susceptibility to the virus. Conclusions Biochemical in vitro assays and transgenic plants generation proved to be useful strategies to assign a role of tomato Twi1 in the plant defence response. Twi1 glycosyltransferase showed to regulate quercetin and kaempferol levels in tomato plants, affecting plant resistance to viral infection.

Tomato glycosyltransferase Twi1 plays a role in flavonoid glycosylation and defence against virus

10.21203/rs.2.10162/v1 ◽

2019 ◽

Author(s):

Laura Campos ◽

María Pilar López-Gresa ◽

Diana Fuertes ◽

José María Bellés ◽

Ismael Rodrigo ◽

...

Keyword(s):

Secondary Metabolites ◽

Higher Plants ◽

Plant Defence ◽

In Vitro Assays ◽

Tomato Plants ◽

Abstract Background: Secondary metabolites play an important role in the plant defensive response. They are produced as a defence mechanism against biotic stress by providing plants with antimicrobial and antioxidant weapons. In higher plants, the majority of secondary metabolites accumulate as glycoconjugates. Glycosylation is one of the commonest modifications of secondary metabolites, and is carried out by enzymes called glycosyltransferases. Results: Here we provide evidence that the previously described tomato wound and pathogen-induced glycosyltransferase Twi1 displays in vitro activity toward the coumarins scopoletin, umbelliferone and esculetin, and the flavonoids quercetin and kaempferol, by uncovering a new role of this gene in plant glycosylation. To test its activity in vivo, Twi1-silenced transgenic tomato plants were generated and infected with Tomato spotted wilt virus. The Twi1-silenced plants showed a differential accumulation of Twi1 substrates and enhanced susceptibility to the virus. Conclusions: Biochemical in vitro assays and transgenic plants generation proved to be useful strategies to assign a role of tomato Twi1 in the plant defence response. Twi1 glycosyltransferase showed to regulate quercetin and kaempferol levels in tomato plants, affecting plant resistance to viral infection.

Plant derived secondary metabolites in cancer therapy: actions, applications, and future prospective of dietaryflavonol (quercetin)

International Research Journal of Multidisciplinary Technovation ◽

10.34256/irjmtcon16 ◽

2019 ◽

Vol 1 (6) ◽

pp. 135-143

Author(s):

Krishnagowdu Saravanan ◽

Jayachandran Halka ◽

Kumaresan Kowsalya ◽

Muthukrishnan Arun

Keyword(s):

Cancer Therapy ◽

Secondary Metabolites ◽

Anticancer Agents ◽

Higher Plants ◽

Lung Tumor ◽

Chemical Constituents ◽

Food Additives ◽

Antioxidant Agents

Higher plants are prominent sources for several bioactive chemical constituents (secondary metabolites) who include photochemical, flavoring agents, fragrant molecules, and food additives. According to WHO estimates, it has been reported that more than 80% of population in developing countries prefer these natural bioactive active compounds for their primary health requirement. At present, conventional chemotherapy is constrained due to the nonselective toxicity to human organs and their usage is limited now a days. In a recent survey, more than 60% of cancer patients have been preferring adjuvant phototherapy along with chemotherapy. Thus, photochemical are being widely used as anticancer agents to target specific pathological pathways underlying cancer with low toxic profiles and side effects. These photochemical are cost-effective and easily accessible to the public to treat cancer diseases. These bioactive photochemical are meticulously belongs to secondary metabolites such as alkaloids, flavonoids, polyphenols.Among them, the flavonoids are polyphenolic substances, which are found in all parts of the plant such as flowers, fruits, leaves, roots, seeds, and bark. They possess high medicinal properties like being anti-cancer, anti-hypertensive, anti-inflammatory, anti-obesity, anti-malarial, antioxidant agents. Quercetin is major flavones associated with a profound antioxidant and medicinal property to prevent the oxidation of lipids in vitro and in vivo, and also exhibits direct proapoptotic effects on tumor cells. This compound has proven efficacy in targeting several cancer cells of breast, colon, prostate, ovarian, and lung tumor in vitro. The present review focuses on the effect of quercetin in cancer therapy.

HIF-1α is a key mediator of the lung inflammatory potential of lithium-ion battery particles

Particle and Fibre Toxicology ◽

10.1186/s12989-019-0319-z ◽

2019 ◽

Vol 16 (1) ◽

Cited By ~ 4

Author(s):

Violaine Sironval ◽

Mihaly Palmai-Pallag ◽

Rita Vanbever ◽

François Huaux ◽

Jorge Mejia ◽

...

Keyword(s):

Inflammatory Responses ◽

Hypoxia Inducible Factor ◽

Lithium Ion ◽

In Vitro Assays ◽

Inhalation Toxicity ◽

Cobalt Nickel ◽

Occupational Settings

Abstract Background Li-ion batteries (LIB) are increasingly used worldwide. They are made of low solubility micrometric particles, implying a potential for inhalation toxicity in occupational settings and possibly for consumers. LiCoO2 (LCO), one of the most used cathode material, induces inflammatory and fibrotic lung responses in mice. LCO also stabilizes hypoxia-inducible factor (HIF) -1α, a factor implicated in inflammation, fibrosis and carcinogenicity. Here, we investigated the role of cobalt, nickel and HIF-1α as determinants of toxicity, and evaluated their predictive value for the lung toxicity of LIB particles in in vitro assays. Results By testing a set of 5 selected LIB particles (LCO, LiNiMnCoO2, LiNiCoAlO2) with different cobalt and nickel contents, we found a positive correlation between their in vivo lung inflammatory activity, and (i) Co and Ni particle content and their bioaccessibility and (ii) the stabilization of HIF-1α in the lung. Inhibition of HIF-1α with chetomin or PX-478 blunted the lung inflammatory response to LCO in mice. In IL-1β deficient mice, HIF-1α was the upstream signal of the inflammatory lung response to LCO. In vitro, the level of HIF-1α stabilization induced by LIB particles in BEAS-2B cells correlated with the intensity of lung inflammation induced by the same particles in vivo. Conclusions We conclude that HIF-1α, stabilized in lung cells by released Co and Ni ions, is a mechanism-based biomarker of lung inflammatory responses induced by LIB particles containing Co/Ni. Documenting the Co/Ni content of LIB particles, their bioaccessibility and their capacity to stabilize HIF-1α in vitro can be used to predict the lung inflammatory potential of LIB particles.

Expression of a Synthesized Gene Encoding Cationic Peptide Cecropin B in Transgenic Tomato Plants Protects against Bacterial Diseases

Applied and Environmental Microbiology ◽

10.1128/aem.00698-09 ◽

2009 ◽

Vol 76 (3) ◽

pp. 769-775 ◽

Cited By ~ 55

Author(s):

Pey-Shynan Jan ◽

Hsu-Yuang Huang ◽

Hueih-Min Chen

Keyword(s):

Xanthomonas Campestris ◽

Plant Pathogens ◽

Transgenic Tomato ◽

Plant Diseases ◽

Gene Encoding ◽

Tomato Plants ◽

Cecropin B ◽

Transgenic Tomato Plants

ABSTRACT The cationic lytic peptide cecropin B (CB), isolated from the giant silk moth (Hyalophora cecropia), has been shown to effectively eliminate Gram-negative and some Gram-positive bacteria. In this study, the effects of chemically synthesized CB on plant pathogens were investigated. The S50s (the peptide concentrations causing 50% survival of a pathogenic bacterium) of CB against two major pathogens of the tomato, Ralstonia solanacearum and Xanthomonas campestris pv. vesicatoria, were 529.6 μg/ml and 0.29 μg/ml, respectively. The CB gene was then fused to the secretory signal peptide (sp) sequence from the barley α-amylase gene, and the new construct, pBI121-spCB, was used for the transformation of tomato plants. Integration of the CB gene into the tomato genome was confirmed by PCR, and its expression was confirmed by Western blot analyses. In vivo studies of the transgenic tomato plant demonstrated significant resistance to bacterial wilt and bacterial spot. The levels of CB expressed in transgenic tomato plants (∼0.05 μg in 50 mg of leaves) were far lower than the S50 determined in vitro. CB transgenic tomatoes could therefore be a new mode of bioprotection against these two plant diseases with significant agricultural applications.

Plastoquinone-9 biosynthesis in cyanobacteria differs from that in plants and involves a novel 4-hydroxybenzoate solanesyltransferase

Biochemical Journal ◽

10.1042/bj20111796 ◽

2012 ◽

Vol 442 (3) ◽

pp. 621-629 ◽

Cited By ~ 14

Author(s):

Radin Sadre ◽

Christian Pfaff ◽

Stephan Buchkremer

Keyword(s):

Biosynthetic Pathway ◽

In Vitro Assays ◽

Transport Chain ◽

In Vivo Labelling ◽

Membrane Bound ◽

Transformation Processes ◽

Synechocystis Sp

PQ-9 (plastoquinone-9) has a central role in energy transformation processes in cyanobacteria by mediating electron transfer in both the photosynthetic as well as the respiratory electron transport chain. The present study provides evidence that the PQ-9 biosynthetic pathway in cyanobacteria differs substantially from that in plants. We identified 4-hydroxybenzoate as being the aromatic precursor for PQ-9 in Synechocystis sp. PCC6803, and in the present paper we report on the role of the membrane-bound 4-hydroxybenzoate solanesyltransferase, Slr0926, in PQ-9 biosynthesis and on the properties of the enzyme. The catalytic activity of Slr0926 was demonstrated by in vivo labelling experiments in Synechocystis sp., complementation studies in an Escherichia coli mutant with a defect in ubiquinone biosynthesis, and in vitro assays using the recombinant as well as the native enzyme. Although Slr0926 was highly specific for the prenyl acceptor substrate 4-hydroxybenzoate, it displayed a broad specificity with regard to the prenyl donor substrate and used not only solanesyl diphosphate, but also a number of shorter-chain prenyl diphosphates. In combination with in silico data, our results indicate that Slr0926 evolved from bacterial 4-hydroxybenzoate prenyltransferases catalysing prenylation in the course of ubiquinone biosynthesis.

L1CAM promotes ovarian cancer stemness and tumor initiation via FGFR1/SRC/STAT3 signaling

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-02117-z ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Marco Giordano ◽

Alessandra Decio ◽

Chiara Battistini ◽

Micol Baronio ◽

Fabrizio Bianchi ◽

...

Keyword(s):

Molecular Mechanisms ◽

Genetic Manipulation ◽

Tumor Formation ◽

In Vitro Assays ◽

Tumor Initiation ◽

Loss Of Function ◽

Stat3 Signaling

Abstract Background Cancer stem cells (CSC) have been implicated in tumor progression. In ovarian carcinoma (OC), CSC drive tumor formation, dissemination and recurrence, as well as drug resistance, thus contributing to the high death-to-incidence ratio of this disease. However, the molecular basis of such a pathogenic role of ovarian CSC (OCSC) has been elucidated only to a limited extent. In this context, the functional contribution of the L1 cell adhesion molecule (L1CAM) to OC stemness remains elusive. Methods The expression of L1CAM was investigated in patient-derived OCSC. The genetic manipulation of L1CAM in OC cells provided gain and loss-of-function models that were then employed in cell biological assays as well as in vivo tumorigenesis experiments to assess the role of L1CAM in OC cell stemness and in OCSC-driven tumor initiation. We applied antibody-mediated neutralization to investigate L1CAM druggability. Biochemical approaches were then combined with functional in vitro assays to study the molecular mechanisms underlying the functional role of L1CAM in OCSC. Results We report that L1CAM is upregulated in patient-derived OCSC. Functional studies showed that L1CAM promotes several stemness-related properties in OC cells, including sphere formation, tumor initiation and chemoresistance. These activities were repressed by an L1CAM-neutralizing antibody, pointing to L1CAM as a druggable target. Mechanistically, L1CAM interacted with and activated fibroblast growth factor receptor-1 (FGFR1), which in turn induced the SRC-mediated activation of STAT3. The inhibition of STAT3 prevented L1CAM-dependent OC stemness and tumor initiation. Conclusions Our study implicate L1CAM in the tumorigenic function of OCSC and point to the L1CAM/FGFR1/SRC/STAT3 signaling pathway as a novel driver of OC stemness. We also provide evidence that targeting this pathway can contribute to OC eradication.

Targeting Myeloid-Cell Specific Integrin α9β1 Inhibits Arterial Thrombosis in Mice

Blood ◽

10.1182/blood-2019-127006 ◽

2019 ◽

Vol 134 (Supplement_1) ◽

pp. 3581-3581

Author(s):

Nirav Dhanesha ◽

Manasa K Nayak ◽

Prakash Doddapattar ◽

Anil K Chauhan

Keyword(s):

Platelet Aggregation ◽

Arterial Thrombosis ◽

Conflicts Of Interest ◽

Myeloid Cell ◽

Cathepsin G ◽

In Vitro Assays ◽

Laser Injury

Background: Coordinated interactions between neutrophils, platelets and endothelial cells contribute towards the development of arterial thrombosis. Neutrophils along with platelets are the first immune cells that are recruited at the site of endothelial activation/injury or infection. Recent studies have suggested that neutrophils modulate thrombosis via several mechanisms, including NETosis (formation of neutrophil extracellular traps). The integrin α9 is highly expressed on neutrophils while platelets do not express it. The integrin α9 up-regulated upon neutrophil activation and is implicated in stable adhesion and transmigration. The mechanisms underlying the role of integrin α9 towards the progression of arterial thrombosis has not been explored yet. Objective: To elucidate the mechanistic insights into the role of myeloid-cell specific integrin α9 in neutrophil adhesion and arterial thrombosis. Methods: We generated novel myeloid-specific α9-/- mice (α9fl/fl LysMcre+l-) by crossing α9fl/fl with LysMcr+/+mice. Littermates α9fl/flLysMcre-l-mice were used as controls. Standardized in vitro assays were used to evaluate the role of integrin α9 in neutrophil mediated platelet aggregation, NETosis and Cathepsin-G release. Susceptibility to arterial thrombosis and hemostasis was evaluated in vivo (FeCl3-induced carotid and laser-injury induced mesenteric artery thrombosis models) by utilizing intravital microscopy and tail bleeding assay respectively. Results: α9fl/flLysMCre+/-mice developed smaller thrombi (~40% occlusion), when compared with α9fl/flmice (~80% occlusion, 10 minutes post-FeCl3 induced injury). The mean time to complete occlusion was significantly prolonged in α9fl/flLysMCre+/-mice (P<0.05 vs α9fl/fl mice). Consistent with this, α9fl/flLysMCre+/-mice displayed significantly decreased platelet mean fluorescence intensity (MFI) and reduced rate of thrombus growth in laser injury-induced thrombosis model (P<0.05 vs. α9fl/fl mice). Together, these results suggest that myeloid cell-specific integrin α9 contributes to the experimental thrombosis at arterial shear rates. Monocytes depletion experiments demonstrated a minimal role for monocyte in progression of arterial thrombosis. In vitro mechanistic studies demonstrated a reduction in neutrophil-mediated platelet aggregation and cathepsin-G secretion in myeloid cell-specific integrin α9-/- mice, when compared with litter-mates control wild-type mice. Notably, the percentage of cells releasing NETs was markedly reduced in myeloid cell-specific integrin α9-/- mice that was concomitant with reduced MPO levels in carotid thrombus of α9fl/flLysMCre+/-mice. Together, these results suggest most likely integrin α9 expressed on neutrophils, but not monocytes, promotes arterial thrombosis. Comparable tail bleeding time between α9fl/flLysMcreand littermate α9fl/fl mice suggested that myeloid-cell specific deficiency of integrin α9 does not alter hemostasis. Conclusion: These findings reveal a novel role for integrin α9 in modulation of arterial thrombosis. While the clinical implications of these findings remains to be explored, we suggest that targeting integrin α9 may reduce post reperfusion thrombo-inflammatory injury, following acute myocardial infarction or stroke. Disclosures No relevant conflicts of interest to declare.