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Biology subjects

Bisht, V.

Publications and source records attributed to Bisht, V..

3 recordsLinked to original sources

A high-resolution atlas of cattle regulatory variants and their cross-species activity in matched human cells

Identifying causal noncoding variants underlying complex traits in cattle remains challenging because high-resolution functional maps of regulatory variation are lacking. Here we combine massively parallel reporter assays with graph genomics to measure autonomous transcriptional activity from >1.5 billion DNA fragments spanning both cattle subspecies. In primary bovine cells, we assay >15 million variants and identify >150,000 expression-modulating variants enriched at cattle eQTL and GWAS loci. This enables the refinement of broad association signals to small sets of candidate functional regulatory variants. Our haplotype-aware framework captures rare, multi-allelic and tightly linked variants poorly resolved by conventional eQTL studies, and quantifies the disproportionate impact of larger variants on transcription. Furthermore, we use these data to train a deep-learning model that successfully predicts bovine promoter activity directly from sequence. Profiling the same cattle DNA in matched primary human cells reveals widespread conservation of promoter and enhancer activity, allelic effects and regulatory grammar, supporting the transfer of annotations and models across species. However, species-dependent effects are enriched in evolutionarily young sequences and p53-family motifs, highlighting the limits to simple cross-species extrapolation. Together, these data provide a high-resolution atlas of cattle regulatory variation and a framework for prioritising causal noncoding variants for cattle trait improvement.

genetics↗

A novel cholesterol dehydrogenase from Limosilactobacillus fermentum catalyzes the direct conversion of cholesterol to coprostanol

On the one hand, cholesterol is a foundational molecule for various structural and biochemical pathways, while elevated cholesterol levels are associated with cardiovascular diseases. Some selected strains of Lactobacilli are also known for modulating cholesterol levels. However, the molecular mechanism of cholesterol transformation by lactobacilli has remained challenging to reveal. This study reports the discovery and role of a microbial 3{beta}-OH-{Delta}5-6-cholesterol-5{beta}-reductase from Limosilactobacillus fermentum NKN51, which directly converts cholesterol to coprostanol, thereby resolving the enigma. Protein engineering of the reductase enzyme identified the cholesterol and NADP+ interacting amino acids, detailing the catalytic mechanism of 5{beta}ChR. Phylogenetic studies emphasize the abundance of 5{beta}ChRs in gut commensal lactobacilli, which shares a common ancestor with plant 5{beta} reductases. Meta-analysis results of healthy participant microbiomes underline the significance of 5{beta}ChR homologs, while a cohort study reveals an association between higher 5{beta}ChR abundance and diabetes. The discovery of the 5{beta}ChR enzyme and its molecular mechanism in cholesterol metabolism pave the way for a better understanding of the gut-associated microbiome and the design of practical applications to ameliorate dyslipidemia.

molecular biology↗

A Live Attenuated Vaccine Model Confers Cross-Protective Immunity Against Different Species of Leptospira spp.

Leptospirosis is the leading zoonotic disease in terms of morbidity and mortality worldwide with a great health and economic impact in both humans and animals. Effective prevention is urgently needed as rapid urbanization, climate change and drivers of disease transmission continue to intensify. The key challenge has been developing a widely-applicable vaccine that protects against the 13 different pathogenic species and >300 serovars that can cause leptospirosis, providing a major public health benefit and opportunity to leverage One Health approaches. Live attenuated mutants that can boost immunity and induce protection are enticing vaccine candidates and poorly explored in the field. We evaluated a recently characterized motility-deficient mutant lacking the expression of a flagellar protein, FcpA. Although the fcpA- mutant has lost its ability to penetrate mucous membranes and cause disease, a transient bacteremia prior to clearance by the host immune response was observed. In two animal models, immunization with a single dose of the fcpA- mutant was sufficient to induce robust anti-protein antibodies response that promoted protection against infection with different Leptospira spp. species. Furthermore, characterization of the immune response identified a small repertoire of biologically relevant proteins that are highly conserved among pathogenic Leptospira species and potential correlates of cross-protective immunity.

microbiology↗