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Yadav, S.

Publications and source records attributed to Yadav, S..

102 records · Page 6Linked to original sources

Rare genera differentiate urban green space soil bacterial communities in three cities across the world

Vegetation complexity is potentially important for urban green space designs aimed at fostering microbial biodiversity to benefit human health. Exposure to urban microbial biodiversity may influence human health outcomes via immune training and regulation. In this context, improving human exposure to microbiota via biodiversity-centric urban green space designs is an underused opportunity. There is currently little knowledge on the association between vegetation complexity (i.e., diversity and structure) and soil microbiota of urban green spaces. Here, we investigated the association between vegetation complexity and soil bacteria in urban green spaces in Bournemouth, UK; Haikou, China; and the City of Playford, Australia by sequencing the 16S rRNA V4 gene region of soil samples and assessing bacterial diversity. We characterized these green spaces as having low or high vegetation complexity and explored whether these two broad categories contained similar bacterial community compositions and diversity around the world. Within cities, we observed significantly different alpha and beta diversities between vegetation complexities; however, these results varied between cities. Rare genera (< 1 % relative abundance individually, on average 35 % relative abundance when pooled) were most likely to be significantly different in sequence abundance between vegetation complexities and therefore explained much of the differences in microbial communities observed. Overall, general associations exist between soil bacterial communities and vegetation complexity, although these are not consistent between cities. Therefore, more in-depth work is required to be done locally to derive practical actions to assist the conservation and restoration of microbial communities in urban areas.

ecology↗

MicroRNA775 and target Galactosyltransferase (GALT9) module regulates recovery from submergence induced senescence by modulating SAGs in Arabidopsis thaliana

Submergence induced hypoxic condition is one of the abiotic stresses which negatively affects the plant growth and development, and causes early onset of senescence. Hypoxic conditions ateres the expression of a number of non-coding microRNAs (miRNAs), besides protein-coding genes. However, the molecular function of stress-induced miRNA in submergence induced physiological or developmental changes and recovery remains to be understood. The expression of miR775 is highly induced under hypoxic stress conditions. Here, we show that miR775 is a potential post-transcriptional regulator number of targets, including Galactosyltransferase (GALT9). The expression of miR775 and target GALT9 was significantly induced and reduced respectively at 24 hours of submergence. The overexpression of miR775 (miR775-Oe) confers enhanced recovery from submergence stress and reduced accumulation of ROS, in contrast to wild type and endogenous target mimic of miR775 (MIM775) Arabidopsis plants. We observed a similar recovery phenotype in case of target galt9 mutant plants, indicating the role of miR775-GALT9 module in recovery from submergence. Further, we showed that the expression of SENESCENCE ASSOCIATED GENES (SAGs), such as SAG12, SAG29, and ORE1. was increased in MIM775 and reduced in miR775-Oe and galt9 plants. Thus, our results suggest that miR775-GALT9 module plays a crucial role in the recovery from submergence by modulating the expression of SAGs through differential accumulation of ROS.

plant biology↗

PIWI silencing mechanism involving the retrotransposon nimbus orchestrates resistance to infection with Schistosoma mansoni in the snail vector, Biomphalaria glabrata

BackgroundSchistosomiasis remains widespread in many regions despite efforts at its elimination. By examining changes in the transcriptome at the host-pathogen interface in the snail Biomphalaria glabrata and the blood fluke Schistosoma mansoni, we previously demonstrated that an early stress response in juvenile snails, manifested by induction of heat shock protein 70 (Hsp 70) and Hsp 90 and of the reverse transcriptase (RT) domain of the B. glabrata non-LTR-retrotransposon, nimbus, were critical for B. glabrata susceptibility to S. mansoni. Subsequently, juvenile B. glabrata BS90 snails, resistant to S. mansoni at 25{degrees}C become susceptible by the F2 generation when maintained at 32{degrees}C, indicating an epigenetic response. Methodology/Principal FindingsTo better understand this plasticity in susceptibility of the BS90 snail, mRNA sequences were examined from S. mansoni exposed juvenile BS90 snails cultured either at 25{degrees}C (permissive temperature) or 32{degrees}C (non-permissive). Comparative analysis of transcriptomes from snails cultured at the non-permissive and permissive temperatures revealed that whereas stress related transcripts dominated the transcriptome of susceptible BS90 juvenile snails at 32{degrees}C, transcripts encoding proteins with a role in epigenetics, such as PIWI (BgPiwi), chromobox protein homolog 1 (BgCBx1), histone acetyl transferase histone deacetylase (HDAC) and metallotransferase (MT) were highly expressed in those cultured at 25{degrees}C. To further determine a role for BgPiwi in B. glabrata susceptibility to S. mansoni, siRNA corresponding to the BgPiwi encoding transcript was utilized to suppress expression of BgPiwi, rendering the resistant BS90 juvenile snail susceptible to infection at 25{degrees}C. Given transposon silencing activity of PIWI as a facet of its role as guardian of the integrity of the genome, we examined the expression of the nimbus RT encoding transcript at 120 min after infection of resistant BS90 piwi-siRNA treated snails. We observed that nimbus RT was upregulated, indicating that modulation of the transcription of the nimbus RT was associated with susceptibility to S. mansoni in BgPiwi- siRNA treated BS90 snails. Furthermore, treatment of susceptible snails with the RT inhibitor lamivudine, before exposure to S. mansoni, blocked S. mansoni infection concurrent with downregulation of the nimbus RT transcript and upregulation of the BgPiwi encoding transcript in the lamivudine-treated, schistosome-exposed susceptible snails. Conclusions and SignificanceThese findings support a role for the interplay of BgPiwi and nimbus in the epigenetic modulation of plasticity of resistance/susceptibility in the snail-schistosome relationship.

molecular biology↗

Exposure of mammary cells to lipid activates gene expression changes associated with ER-negative breast cancer via chromatin remodeling.

Improved understanding of local breast biology that favors the development of estrogen receptor negative (ER-) breast cancer (BC) would foster better prevention strategies. We have previously shown that overexpression of specific lipid metabolism genes is associated with the development of ER- BC. We now report results of exposure of MCF-10A cells and mammary organoids to representative medium- and long-chain polyunsaturated fatty acids. This exposure caused a dynamic and profound change in gene expression, accompanied by changes in chromatin packing density, chromatin accessibility and histone posttranslational modifications (PTMs). We identified 38 metabolic reactions that showed significantly increased activity, including reactions related to one-carbon metabolism. Among these reactions are those that produce S-adenosyl-L-methionine for histone PTMs. Utilizing both an in-vitro model and samples from women at high risk for ER- BC, we show that lipid exposure engenders gene expression, signaling pathway activation, and histone marks associated with the development of ER- BC.

cancer biology↗

Desert Dingo (Canis lupus dingo) genome provides insights into their role in the Australian ecosystem.

The dingo is Australias iconic top-order predator and arrived on the continent between 5,000-8,000 years ago. To provide an unbiased insight into its evolutionary affiliations and biological interactions, we coupled long-read DNA sequencing with a multiplatform scaffolding approach to produce an ab initio genome assembly of the desert dingo (85X coverage) we call CanLup_DDS. We compared this genome to the Boxer (CanFam3.1) and German Shepherd dog (CanFam_GSD) assemblies and characterized lineage-specific and shared genetic variation ranging from single- to megabase pair-sized variants. We identified 21,483 dingo-specific and 16,595 domestic dog-specific homozygous structural variants mediating genic and putative regulatory changes. Comparisons between the dingo and domestic dog builds detected unique inversions on Chromosome 16, structural variations in genes linked with starch metabolism, and seven differentially methylated genes. To experimentally assess genomic differences 17 dingoes and 15 German Shepherd dogs were fed parallel diets for 14 days. In dingoes, low AMY2B copy number and serum amylase levels are linked with high cholesterol and LDL levels. Gut microbiome analyses revealed enrichment of the family Clostridiaceae, which can utilize complex resistant starch, while scat metabolome studies identified high phenylethyl alcohol concentrations that we posit are linked with territory marking. Our study provides compelling genomic, microbiome, and metabolomic links showing the dingo has distinct physiology from domestic breed dogs with a unique role in the ecosystem.

genomics↗

Metabolomics shows the Australian dingo has a unique plasma profile

Dingoes have not been artificially selected in the past 3,500 years. They occupy a wide range of the Australian mainland and play a crucial role as an apex predator with a generalist omnivorous feeding behaviour. In contrast, humans have selected breed dogs for novel and desirable traits. First, we explore whether the distinct evolutionary histories of dingoes and domestic dogs can lead to plasma metabolomic differences. We study metabolite composition differences between dingoes (n=15) and two domestic dog breeds (Basenji n= 9 and German Shepherd Dog: GSD n=10). After accounting for within group variation, 62 significant metabolite differences were detected between dingoes and domestic dogs, with a greater number of differences in protein (n= 14) and lipid metabolites (n= 12). Most differences were observed between dingoes and domestic dogs and fewest between the domestic dog breeds. Second, we investigate variation between pure dingoes (n=10) and dingo-dog hybrids (n=10) as hybridisation is common. We detected no significant differences in metabolite levels between dingoes and dingo-dog hybrids after Bonferroni correction. However, power analyses reported that increasing the sample size to 15 could result in differences in uridine 5-diphosphogalactose (UDPgal) levels related to galactose metabolism. We suggest this may be related to an increase in Amylase 2B copy number in hybrids. Our study illustrates that the dingo metabolome is significantly different from domestic dog breeds and hybridisation is likely to influence carbohydrate metabolism.

ecology↗

Precision design of single and multi-heme de novo proteins

The de novo design of simplified porphyrin-binding helical bundles is a versatile approach for the construction of valuable biomolecular tools to both understand and enhance protein functions such as electron transfer, oxygen binding and catalysis. However, the methods utilised to design such proteins by packing hydrophobic side chains into a buried binding pocket for ligands such as heme have typically created highly flexible, molten globule-like structures, which are not amenable to structural determination, hindering precise engineering of subsequent designs. Here we report the crystal structure of a de novo two-heme binding "maquette" protein, 4D2, derived from the previously designed D2 peptide, offering new opportunities for computational design and re-engineering. The 4D2 structure was used as a basis to create a range of heme binding proteins which retain the architecture and stability of the initial crystal structure. A well-structured single-heme binding variant was constructed by computational sequence redesign of the hydrophobic protein core, assessed by NMR, and utilised for experimental validation of computational redox prediction and design. The structure was also extended into a four-heme binding helical bundle resembling a molecular wire. Despite a molecular weight of only 24kDa, imaging by CryoEM illustrated a remarkable level of detail in this structure, indicating the positioning of both the secondary structure and the heme cofactors. The design and determination of atomic-level resolution in such de novo proteins is an invaluable resource for the continued development of novel and functional protein tools.

synthetic biology↗

Breaking a Strong Amide Bond: Structure and Properties of Dimethylformamidase.

Dimethylformamidase (DMFase) breaks down the human-made synthetic solvent N,N-dimethyl formamide(DMF) used extensively in industry(1). DMF is not known to exist in nature and was first synthesized in 1893. In spite of the recent origin of DMF, certain bacterial species such as Paracoccus, Pseudomonas, and Alcaligenes have evolved pathways to breakdown DMF and use them as carbon and nitrogen source for growth(2, 3). The work presented here provides a molecular basis for the ability of DMFase from Paracoccus to function in exacting conditions of high solvent concentrations, temperature and ionic strength to catalyze the hydrolysis of a stable amide bond. The structure reveals a multimeric complex of the 2{beta}2 type or (2{beta}2)2 type. One of the three domains of the large subunit and the small subunit are hitherto undescribed folds and as yet of unknown evolutionary origin. The active site is made of a distinctive mononuclear iron that is coordinated by two tyrosine residues and a glutamic acid residue. The hydrolytic cleavage of the amide bond is catalyzed at the Fe3+ site with a proximal glutamate probably acting as the base. The change in the quaternary structure is salt dependent with high salt resulting in the larger oligomeric state. Kinetic characterization reveals an enzyme that shows cooperativity between subunits and the structure provides clues on the interconnection between the active sites. Significance StatementN,N-dimethyl formamide(DMF) is a commonly used industrial solvent that was first synthesized in 1893. The properties that make DMF a highly desired solvent also makes it a difficult compound to breakdown. Yet, certain bacteria have evolved to survive in environments polluted by DMF and have enzymes that breakdown DMF and use it as their carbon and nitrogen source. The molecular structure of the enzyme that breaks down the stable amide bond in these bacteria, reveals two new protein folds and a unique mononuclear iron active site. The work reported here provides the structural and biochemical framework to query the evolutionary origins of the protein, as well as in engineering this enzyme for use in bioremediation of a human made toxic solvent.

biochemistry↗

Local auxin biosynthesis promotes stem cell differentiation and organogenesis in Arabidopsis shoot apex

Shoot apical meristem (SAM) of higher plants is comprises of three distinct functional zones. Central zone (CZ) is placed at the meristem summit and harbors pluripotent stem cells. Stem cells undergoes cell division within the CZ and give rise descendants which enters into the surrounding peripheral zone (PZ), where they get recruited into organs. Stem cell daughters that get pushed underneath the CZ form rib meristem (RM). RM cells differentiate into stem tissue and vascular bundles. Understanding how stem cell daughters differentiate into PZ and RM cell types is essential to unravel the mechanism of meristem development in higher plants. Here, we show that meristem patterning and lateral organ primordia formation, besides intercellular transport, are also regulated by auxin biosynthesis mediated by two closely related genes belonging to TRYPTOPHAN AMINOTRANSFERASE family. In Arabidopsis SAM, TAA1 and TAR2 are required to maintain auxin responses and identity of PZ cell types. Furthermore, our genetic analysis shows that in the absence of local auxin production and transport differentiation of stem cells into PZ and RM cell types is stalled causing a complete arrest of shoot growth and development. Our study revealed that auxin biosynthesis and transport together control the patterning of SAM into PZ and RM cell types.

plant biology↗

Prediction of the functional impact of missense variants in BRCA1 and BRCA2 with BRCA-ML

In silico predictions of missense variants is an important consideration when interpreting variants of uncertain significance (VUS) in the BRCA1 and BRCA2 genes. We trained and evaluated hundreds of machine learning algorithms based on results from validated functional assays to better predict missense variants in these genes as damaging or neutral. This new optimal \"BRCA-ML\" model yielded a substantially more accurate method than current algorithms for interpreting the functional impact of variants in these genes, making BRCA-ML a valuable addition to data sources for VUS classification.

bioinformatics↗

Altered gut microbiota and immunity defines Plasmodium vivax survival in Anopheles stephensi

Blood feeding-enriched gut-microbiota boosts mosquitoes anti-Plasmodium immunity. Here, we ask how Plasmodium vivax alters microbiota, anti-Plasmodial immunity and impact tripartite Plasmodium-mosquito-microbiota interactions in the gut lumen. Using a metagenomics analysis, we predominantly detect Elizabethkingia meningitis and Pseudomonas sps. in naive mosquitoes. Naive blood fed gut shows a heightened presence of Elizabethkingia, Pseudomonas and Serratia. A parallel RNAseq analysis of blood-fed midguts identify Elizabethkingia-transcripts, which may have role in iron metabolism. Post, a Plasmodium vivax infected blood-meal, however, we do not detect bacterial until circa 36 hours. Intriguingly, transcriptional expression of a selected array of antimicrobial arsenal cecropins 1-2, defensin-1 and gambicin remains low during the first 36 hours-a time frame when ookinietes/early oocysts invade gut. We conclude during the preinvasive phase, Plasmodium vivax outcompetes midgut-microbiota. Suppression of important immune factors, likely due to altered microbiota, may enhance Plasmodium vivax survival. Additional finding of a novel Wolbachia association warrants further research to design paratransgenesis tools for malaria control.\n\nAuthor SummarySuccessful malaria transmission relies on the competitive interactions of Plasmodium and mosquitos tissue specific immune potential. Within 24hrs of blood meal gut-microbiota grows exponentially and lead to robust enhancement of mosquito immune response, which is detrimental to parasite survival and development. But the mechanism how Plasmodium manages to evade this pre-invasive immune barrier is not well known. We investigated the influence of tripartite gut-microbiome-parasite interaction on human malaria parasite Plasmodium vivax in its natural/native vector Anopheles stephensi. Surprisingly we found that infectious blood meal lead to dramatic suppression in gut-bacteria population, a plausible strategy of P. vivax ookinetes to avoid immune responses. Our study suggests that for its own survival Plasmodium vivax causes early suppression of bacterial population, possibly by scavenging Fe from the blood meal which is indispensable for bacterial growth. Disruption and manipulation of this gut-microbe-interaction may help to design new paratransgenesis molecular tool for malaria control.

genomics↗

Serotonergic and catecholaminergic (dopaminergic) oscillations in the reproductive regulation of Japanese quail

Specific temporal phase relation of serotonergic and dopaminergic oscillations alters reproductive responses in many species. Aim of the study was to confirm whether effect of serotonergic drug (5-HTP) and dopaminergic drug (L-DOPA) is due to their conversion into serotonin and dopamine respectively or other products. For this study, PCPA (p-chlorophenylalanine, a long lasting inhibitor of serotonin synthesis), DDC (Diethyldithiocarbamate, which inhibits biosynthesis of nor-adrenaline), -MT (Methyl-p-tyrosine, an inhibitor for the conversion of tyrosine to DOPA) and DOPS (Dihydroxyphenylserine, a specific precursor for noradrenaline) were used in different groups in addition to 5-HTP and L-DOPA given at specific time interval. Reproductive responses monitored at 10 weeks post treatment indicate that gonadal activity was significantly low in HTP:DOPA (8-hr quail), HTP+PCPA:DOPA and HTP:DOPA+DDC quail compare to control (S:S). However, gonadal activity of HTP:S(HTP control), S:DOPA(DOPA control) and HTP: -MT+DOPS was not different from S:S control and remained in active condition. These findings indicate that it is not the dose of neurotransmitter precursor drugs (5-HTP and L-DOPA) and the neurotransmitters (serotonin and dopamine itself) that cause the effect, instead it is the function of interval between the drug administration which induces or entrains specific phase relation between serotonergic and dopaminergic oscillations. Further, gonadal suppression observed in HTP:DOPA, HTP+PCPA:DOPA and HTP:DOPA+DDC group three groups is not due to injection of 5-HTP or L-DOPA (alone) but due to conversion of administered 5-HTP into serotonin and conversion of L-DOPA (administered) into dopamine; not due to their further conversion into catecholamines other than dopamine i.e. noradrenaline or adrenaline.

animal behavior and cognition↗