sli-3 negatively regulates the LET-23/epidermal growth factor receptor-mediated vulval induction pathway in Caenorhabditis elegans

The LIN-3-LET-23-mediated inductive signaling pathway plays a major role during vulval development in C. elegans. Studies on the components of this pathway have revealed positive as well as negative regulators that function to modulate the strength and specificity of the signal transduction cascade. We have carried out genetic screens to identify new regulators of this pathway by screening for suppressors of lin-3 vulvaless phenotype. The screens recovered three loci including alleles of gap-1 and a new gene represented by sli-3. Our genetic epistasis experiments suggest that sli-3 functions either downstream or in parallel to nuclear factors lin-1 and sur-2. sli-3 synergistically interacts with the previously identified negative regulators of the let-23 signaling pathway and causes excessive cell proliferation. However, in the absence of any other mutation sli-3 mutant animals display wild-type vulval induction and morphology. We propose that sli-3 functions as a negative regulator of vulval induction and defines a branch of the inductive signaling pathway. We provide evidence that sli-3 interacts with the EGF signaling pathway components during vulval induction but not during viability and ovulation processes. Thus, sli-3 helps define specificity of the EGF signaling to induce the vulva.     

Genetic dissection of days to flowering via genome-wide association studies in Turkish common bean germplasm

Common bean is a nutrient‐dense legume crop serving as a source of food for millions of people. Characterization of unexplored common bean germplasm to unlock the phenotypic and genetic variations is still needed to explore the breeding potential of this crop. The current study aimed to dissect the genetic basis having association for days to flowering (DF). A total of 188 common bean accessions collected from 19 provinces of Turkey were used as plant material under five environments and two locations. Analysis of variance (ANOVA) revealed that genotypes and genotype by environment interaction have significant effects on DF. A total of 10 most stable accessions were evaluated from stability analysis. Overall maximum (75) and minimum (54) DF were observed for Hakkari-51 and Mus-46 accessions, respectively. The implemented constellation plot divided studied germplasm according to their DF and growth habit. A total of 7900 DArTseq markers were used for association analysis. Mixed linear model using the Q + K Model resulted a total of 18 DArTseq markers from five environments. DArT-8668385 marker identified in Bolu during 2016 was also associated with DF in Sivas during 2017. Combined data of five years resulted a total of four markers (DArT-22346534, DArT-3369768, DArT-3374613, and DArT-3370801) having significant association ( p  <  0.01 ) for DF. DArT-22346534 present on Pv 08 accounted a maximum of 9.89% variation to the studied trait. A total of four putative candidate genes were predicted from sequences reflecting homology to identified four DArTseq markers. We envisage that exploitation of identified DArTseq markers will hopefully beneficial for the development of new common bean varieties having better adaptation ability to changing climatic conditions.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01029-8.     

Involvement of p53 in gemcitabine mediated cytotoxicity and radiosensitivity in breast cancer cell lines

Gemcitabine (2',2'-difluoro-2'-deoxycytidine; dFdCyd) is one of the anti-metabolites drugs that target DNA replication. We evaluated dFdCyd cytotoxicity and its radiosensitizing ability in human breast cancer cell lines, MCF-7 (wild-type p53) and MDA-MB-231 (mutant-type p53) along with normal mammary epithelial cell line (MCF-12) for comparison. Radiosensitivity and cytotoxicity were measured by the clonogenic survival assays. DNA DSBs was studied by Pulse Field Gel Electrophoresis (PFGE) and cell cycle distribution was analyzed by flow cytometry. MDA-MB-231 cells were the most sensitive to the cytotoxicity of dFdCyd (IC(50) 5 nM) then MCF-7 (IC(50) 10nM), whereas MCF-12 cells were the most resistant to the cytotoxicity of dFdCyd (IC(50) 70 nM). MCF-12 and MCF-7 cell lines did not show any radiosensitization to dFdCyd, whereas the MDA-MB-231 cells showed significantly increased radioresistant to dFdCyd at equimolar concentration (p=0.002) and at IC(50) concentration (p<0.001). The DNA double strand breaks (DSBs) repair showed that dFdCyd neither increases DNA DSBs nor decreases the rate of their repair in MCF-12 and MCF-7 cell lines, while the same treatment in MDA-MB-231 cell line led to decrease the rate of DSBs or increase the rate of DNA repair (p=0.034). Therefore, dFdCyd is a cytotoxic agent, especially in the cancer cells irrespective of having wild-type or mutated p53 protein, but it is not effective as radiosensitizer in the cell lines used in this study. dFdCyd combined with radiation reduces the efficacy of chemo-radiotherapy in p53 mutated cells. Therefore, p53-mutated cancer could be a counter-indication for radiation-gemcitabine combined treatment.     

A biphasic endothelial stress-survival mechanism regulates the cellular response to vascular endothelial growth factor A

Vascular endothelial growth factor A (VEGF-A) is an essential cytokine that regulates endothelial function and angiogenesis. VEGF-A binding to endothelial receptor tyrosine kinases such as VEGFR1 and VEGFR2 triggers cellular responses including survival, proliferation and new blood vessel sprouting. Increased levels of a soluble VEGFR1 splice variant (sFlt-1) correlate with endothelial dysfunction in pathologies such as pre-eclampsia; however the cellular mechanism(s) underlying the regulation and function of sFlt-1 are unclear. Here, we demonstrate the existence of a biphasic stress response in endothelial cells, using serum deprivation as a model of endothelial dysfunction. The early phase is characterized by a high VEGFR2:sFlt-1 ratio, which is reversed in the late phase. A functional consequence is a short-term increase in VEGF-A-stimulated intracellular signaling. In the late phase, sFlt-1 is secreted and deposited at the extracellular matrix. We hypothesized that under stress, increased endothelial sFlt-1 levels reduce VEGF-A bioavailability: VEGF-A treatment induces sFlt-1 expression at the cell surface and VEGF-A silencing inhibits sFlt-1 anchorage to the extracellular matrix. Treatment with recombinant sFlt-1 inhibits VEGF-A-stimulated in vitro angiogenesis and sFlt-1 silencing enhances this process. In this response, increased VEGFR2 levels are regulated by the phosphatidylinositol-3-kinase and PKB/Akt signaling pathways and increased sFlt-1 levels by the ERK1/2 signaling pathway. We conclude that during serum withdrawal, cellular sensing of environmental stress modulates sFlt-1 and VEGFR2 levels, regulating VEGF-A bioavailability and ensuring cell survival takes precedence over cell proliferation and migration. These findings may underpin an important mechanism contributing to endothelial dysfunction in pathological states.     

Structural insights into the allosteric effects of 4EBP1 on the eukaryotic translation initiation factor eIF4E

The eukaryotic translation initiation factor eIF4E plays key roles in cap-dependent translation and mRNA export. These functions rely on binding the 7-methyl-guanosine moiety (5'cap) on the 5'-end of all mRNAs. eIF4E is regulated by proteins such as eIF4G and eIF4E binding proteins (4EBPs) that bind the dorsal surface of eIF4E, distal to the cap binding site, and modulate cap binding activity. Both proteins increase the affinity of eIF4E for 5'cap. Our understanding of the allosteric effects and structural underpinnings of 4EBP1 or eIF4G binding can be advanced by obtaining structural data on cap-free eIF4E bound to one of these proteins. Here, we report the crystal structure of apo-eIF4E and cap-free eIF4E in complex with a 4EBP1 peptide. We also monitored 4EBP1 binding to cap-free eIF4E in solution using NMR. Together, these studies suggest that 4EBP1 transforms eIF4E into a cap-receptive state. NMR methods were also used to compare the allosteric routes activated by 4EBP1, eIF4G, and the arenavirus Z protein, a negative regulator of cap binding. We observed chemical shift perturbation at the dorsal binding site leading to alterations in the core of the protein, which were ultimately communicated to the unoccupied cap binding site of eIF4E. There were notable similarities between the routes taken by 4EBP1 and eIF4G and differences from the negative regulator Z. Thus, binding of 4EBP1 or eIF4G allosterically drives alterations throughout the protein that increase the affinity of eIF4E for the 5'cap.     

Genetic variants in interferon gamma (<Emphasis Type="Italic">IFN-γ</Emphasis>) gene are associated with resistance against ticks in <Emphasis Type="Italic">Bos taurus</Emphasis> and <Emphasis Type="Italic">Bos indicus</Emphasis>

In tropical and subtropical regions of the world, parasitic diseases are major obstacle in the health and ultimately overall performance of animals. Cattle express heritable, contrasting phenotypes when exposed to ticks depicting genetic nature of trait. IFN-γ is one of the most reported genes critical for innate and adaptive immunity against viral and intracellular infections. To identify its role in resistance for ectoparasite especially tick, genetic characterization of this gene was done in resistant and susceptible animals of Sahiwal cattle (n = 95) and Friesian (n = 92). Nine Polymorphisms were identified, three of them were found in exonic region. One out of nine variants was being reported previously (ss82716193) and was confirmed in Pakistani Sahiwal cattle population as well. Single site analysis of each variant depicted their significance in tick resistant and tick susceptible groups (P < 0.05). The associations using haplotypes were more informative than for single markers. Eighteen different haplotypes resulting from nine polymorphic sites were used in construction of maximum parsimony tree which categorized resistant and susceptible animals in two clades. Genetic markers identified in this study can be useful in future breeding selection programs against tick resistance.     

Exogenous salicylic acid stimulates physiological and biochemical changes to improve growth, yield and active constituents of fennel essential oil

Fennel (Foeniculum vulgare Mill) is a high-value medicinal and essential oil bearing plant used extensively in pharmaceutical, food and cosmetic industries. A pot experiment was carried out in the natural conditions of net house to resolve whether the foliar application of salicylic acid (SA) might enhance the growth, yield and essential oil production of fennel. Plants were sprayed three times with SA. The first spray was carried out at 40?days after sowing (DAS); the second and third sprays were applied one and 2?weeks later, the plants were sprayed with deionised water (control) and different concentrations of SA (10^?5, 10^?4 and 10^?3?M). The foliar spray of SA at 10^?4?M significantly enhanced the vegetative growth (shoot and root lengths, fresh and dry weights), physiological and biochemical characteristics (chl ??a??, chl ??b??, total chlorophyll and carotenoids contents, nitrate reductase activity, carbonic anhydrase activity, leaf-N, -P and -K contents), yield characteristics (number of umbels and fruits, 1,000-seed weight and seed yield) and essential oil yield of fennel. GLC analysis revealed the significant increase in the components of essential oil, viz. trans-anethole (80.4?C84.7?%), methyl chavicol (2.3?C2.5?%) and fenchone (5.6?C7.9?%). It was concluded that foliar spray of SA at 10^?4?M might be employed for enhancing the plant growth as well as yield and quality of essential oil of fennel.     

Plant Growth-Promoting Rhizobacteria Associated with Ancient Clones of Creosote Bush (Larrea tridentata)

Plant growth-promoting rhizobacteria (PGPR) are common components of the rhizosphere, but their role in adaptation of plants to extreme environments is not yet understood. Here, we examined rhizobacteria associated with ancient clones of Larrea tridentata in the Mohave desert, including the 11,700-year-old King Clone, which is oldest known specimen of this species. Analysis of unculturable and culturable bacterial community by PCR-DGGE revealed taxa that have previously been described on agricultural plants. These taxa included species of Proteobacteria, Bacteroidetes, and Firmicutes that commonly carry traits associated with plant growth promotion, including genes encoding aminocyclopropane carboxylate deaminase and β–propeller phytase. The PGPR activities of three representative isolates from L. tridentata were further confirmed using cucumber plants to screen for plant growth promotion. This study provides an intriguing first view of the mutualistic bacteria that are associated with some of the world’s oldest living plants and suggests that PGPR likely contribute to the adaptation of L. tridentata and other plant species to harsh environmental conditions in desert habitats.     

Characterization of 1064 nm Nd:YAG laser-produced cu plasma

The plasma was produced by focusing Nd:YAG laser pulses of 1064 nm wavelength on to a copper target at laser fluences of 5.35, 6.95, and 9.33 J/cm². An ion collector placed along the target surface normal was used to record the time-of-flight (TOF) ion signal during plasma expansion in vacuum. The TOF ion pulses were deconvoluted using the Coulomb-Boltzmann-shifted function to estimate the available Cu ion charge states, equivalent plasma ion temperature, and accelerating potential in the nonequilibrium plasma. The maximum available ion charge state, equivalent plasma ion temperature, and accelerating potential are found to increase with laser fluence. In the local thermal equilibrium conditions, the accelerating potential can be supposed to apply across a distance of the order of the Debye length. The Debye length and, hence, the electric field in the laser produced plasma at three laser fluences values were estimated. The electric field was in the range of 1 MV/cm and increased with laser fluence. In the laser fluence range used in this work, the sum of thermal and adiabatic energy of the ion was slightly higher than its Coulomb energy.