bioRxiv Science⌕ Search

Biology subjects

Mansoor, S.

Publications and source records attributed to Mansoor, S..

12 recordsLinked to original sources

Spatiotemporal graph neural networks reveal conformational binding signature in protein dynamics

Biomolecular function is often controlled by structural and dynamical adaptations to binding events. Although molecular dynamics (MD) simulations can capture these events at atomic resolution, separating functional signatures from stochastic noise remains challenging. Traditional methods often struggle to isolate mechanistically relevant differences across independent replicas. Here, we introduce an explainable deep learning approach that learns state-specific dynamic signatures directly from MD trajectories. By coupling a dynamic protein graph representation with group-aware contrastive learning across independent replicas, the model detects the signatures, filtering out trajectory-specific correlations. An explainable AI framework then maps the identified differences on individual residues. We demonstrate this approach by identifying "binding-ready" conformations in a T4-Lysozyme mutant, recovering the allosteric determinants of peptide recognition in the PDZ3 domain, and isolating a ligand-independent activation signature for the A2A receptor. Our GISTnet-MD method generalizes across unseen data during comparative MD analysis, translating raw trajectory differences into residue-level determinants of protein function.

biophysics↗

A comprehensive water buffalo pangenome reveals extensive structural variation linked to population specific signatures of selection

Water buffalo is a cornerstone livestock species in many low- and middle-income countries, yet major gaps persist in its genomic characterization--complicated by the divergent karyotypes of its two sub-species (swamp and river). Such genomic complexity makes water buffalo a particularly good candidate for the use of graph genomics, which can capture variation missed by linear reference approaches. However, the utility of this approach to improve water buffalo has been largely unexplored. We present a comprehensive pangenome that integrates four newly generated, highly contiguous assemblies of Pakistani river buffalo with available assemblies from both sub- species. This doubles the number of accessible high-quality river buffalo genomes and provides the most contiguous assemblies for the sub-species to date. Using the pangenome to assay variation across 711 global samples, we uncovered extensive genomic diversity, including thousands of large structural variants absent from the reference genome, spanning over 140 Mb of additional sequence. We demonstrate the utility of these data by identifying putative functional indels and structural variants linked to selective sweeps in key genes involved in productivity and immune response across 26 populations. This study represents one of the first successful applications of graph genomics in water buffalo and offers valuable insights into how integrating assemblies can transform analyses of water buffalo and other species with complex evolutionary histories. We anticipate that these assemblies, and the pangenome and putative functional structural variants we have released, will accelerate efforts to unlock water buffalos genetic potential, improving productivity and resilience in this economically important species.

genomics↗

Paroxysmal Slow Wave Events as a diagnostic and predictive biomarker for post-traumatic epilepsy

Traumatic brain injury (TBI) is a major global health concern, affecting more than 40 million people annually. While most cases are mild and present with light symptoms, repeated mild injuries can result in delayed brain pathologies, including cognitive decline, neuropsychiatric complications, and post-traumatic epilepsy (PTE). PTE refers to recurring, unprovoked seizures occurring at least one week after TBI. While the link between moderate to severe TBI and PTE is well established, the epileptogenesis after repetitive mild TBI (rmTBI) is seldom studied. Currently, there are no biomarkers to identify those at risk of developing PTE, and its diagnosis is challenging. Here, we used a rat model to study PTE following rmTBI and assessed human EEG data to identify potential biomarkers for PTE. We employed a closed head TBI model to induce rmTBI, and recorded brain activity using electrocorticography (ECoG) between 2- and 6-months post-injury. Behavioral assessments and post-mortem analysis were also conducted. In humans, we analyzed EEG recordings from the Temple University database to investigate the potential of EEG-derived features for diagnosing PTE. At 6 months post injury, 70% of rmTBI animals developed PTE, compared to 22% in the control group (P=0.01). While neurological assessments following injury did not predict PTE, paroxysmal slow wave events (PSWEs) were found to be a reliable biomarker for PTE prediction. In humans, the percentage time in PSWEs was significantly elevated in PTE patients with epileptiform activity. In conclusion, we suggest PSWEs as a non-invasive, cost-effective biomarker for PTE in rodents and human patients.

neuroscience↗

Comparative Analysis, Diversification and Functional Validation of Plant Nucleotide-Binding Site Domain Genes

Nucleotide-binding site (NBS) domain genes are one of a superfamily of resistance genes involved in plant responses to pathogens. The current study identified presumably identified 12,820 NBS-containing genes across 34 species covering from mosses to monocots and dicots. These identified genes classified into 168 classes with several novel domain architectures patterns encompassing significant diversity among plant species. Several classical (NBS, NBS-LRR, TIR-NBS, TIR-NBS-LRR etc.) and species-specific structural patterns (TIR-NBS-TIR-Cupin_1-Cupin_1, TIR-NBS-Prenyltransf, Sugar_tr-NBS etc.) were discovered. We observed 603 orthogroups (OGs) with some core (most common orthogroups; OG0, OG1, OG2 etc.) and unique (highly specific to species; OG80, OG82 etc) OGs with tandem duplications. The expression profiling presented the putative upregulation of OG2, OG6, and OG15 in different tissues under various biotic and abiotic stresses in susceptible and tolerant plants to CLCuD. The genetic variation between susceptible (Coker 312) and tolerant (Mac7) G. hirsutum accessions identified several unique variants in NBS genes of Mac7 (6,583 varaints) and Coker312 (5,173 variants). The protein-ligand and proteins-protein interaction showed a strong interaction of some putative NBS proteins with ADP/ATP and different core proteins of cotton leaf curl disease virus. The silencing of GaNBS (OG2) in resistant cotton through virus-induced gene silencing (VIGS) demonstrated its putative role in virus tittering. The presented study will be further helpful to understand the plant adaptation mechanism.

plant biology↗

Protein Ensemble Generation through VariationalAutoencoder Latent Space Sampling

Mapping the ensemble of protein conformations that contribute to function and can be targeted by small molecule drugs remains an outstanding challenge. Here we explore the use of soft-introspective variational autoencoders for reducing the challenge of dimensionality in the protein structure ensemble generation problem. We convert high-dimensional protein structural data into a continuous, low-dimensional representation, carry out search in this space guided by a structure quality metric, then use RoseTTAFold to generate 3D structures. We use this approach to generate ensembles for the cancer relevant protein K-Ras, training the VAE on a subset of the available K-Ras crystal structures and MD simulation snapshots, and assessing the extent of sampling close to crystal structures withheld from training. We find that our latent space sampling procedure rapidly generates ensembles with high structural quality and is able to sample within 1 angstrom of held out crystal structures, with a consistency higher than MD simulation or AlphaFold2 prediction. The sampled structures sufficiently recapitulate the cryptic pockets in the held-out K-Ras structures to allow for small molecule docking.

biochemistry↗

Accurate Mutation Effect Prediction using RoseTTAFold

Predicting the effects of mutations on protein function is an outstanding challenge. Here we assess the performance of the deep learning based RoseTTAFold structure prediction and design method for unsupervised mutation effect prediction. Using RoseTTAFold in inference mode, without any additional training, we obtain state of the art accuracy on predicting mutation effects for a set of diverse protein families. Thus, although the architecture of RoseTTAFold was developed to address the protein structure prediction problem, during model training RoseTTAFold acquired an understanding of the mutational landscapes of proteins comparable to that of large recently developed language models. The ability to reason over structure as well as sequence could enable even more precise mutation effect predictions following supervised training.

biochemistry↗

Hexokinase 1b is a novel target for non-small-cell lung cancer

Deregulation of glycolysis is common in non-small cell lung cancer (NSCLC). Hexokinase (HK) enzymes catalyze the phosphoryl-group-transfer in glucose metabolism. There are a very few studies that have begun to reveal the connections between glucose metabolism and splicing programs. Unlike HK2 gene, which is expressed as a single transcript, there are several transcripts of the HK1 gene due to alternative splicing. However, the functional differential roles of HK1 isoforms in glucose metabolism and tumor progression are still elusive. Here, we show that primary NSCLC patient tumor cells metabolically differ from the normal lung epithelium where they display predominant expression of one of the HK1 transcripts, hexokinase1b (HK1b). We utilized CRISPR-Cas9 system to selectively target specific HK1b isoform in NSCLC and show that silencing HK1b in NSCLC cells inhibits tumorigenesis through diminishing glycolysis and proliferation. Our findings constitute the first demonstration of the first biochemical distinction between the HK1 splice variants. Finally, HK1b deletion sensitizes NSCLC cells to standard-of-care, cisplatin treatment, and the combination therapy synergistically increases both apoptotic cell death by cisplatin and autophagic cell death by increased formation of LC3-II associated autophagic vesicles and myelinoid bodies. Notably, loss of HK1b leads to cellular DNA damage, further combination with cisplatin therapy showed significantly increased levels of DNA damage. Importantly, we showed that glycolysis and cisplatin resistance can be restored by adding-back HK1b in HK1b knock-out cells. Our findings reveal that targeting HK1b isoform alone or in combination with cisplatin may represent a novel strategy for NSCLC patients.

cancer biology↗

Fully Transient CRISPR/Cas12f system in plants capable of broad-spectrum resistance against Begomovirus

CRISPR/Cas system has emerged as the most efficient genome editing technology for both eukaryotic and prokaryotic cells. Recently, biologist have started using CRISPR/Cas9 as a defence machinery in plants against DNA viruses by targeting conserve regions of their genome. Considerable resistance requires formation of stable transgenic lines with multiple gRNAs, targeting specific viruses. Development of such transgenic plants is not only time consuming but also there will always be some uncertainty of their efficiency and off targets in plant genome. Newly discovered miniature CRISPR/Cas12f (Cas14a) system has unique ability to chop down every ssDNA fragment once activated through targeted cleavage. Here we report first fully functional transient CRISPR/Cas12f system in plants. We also show that Cas12f with just one gRNA is enough for substantial broad-spectrum resistance against Gemini viruses. Plant phenotype showed nearly complete recovery and qPCR results showed multifold decrease infection of CLCuV in CRISPR/Cas12f treated plants compare to the infected plants (infected with CLCuMuV and CLCuKV). Smaller size, broad range and more efficiency make Cas12f a superior alternative of Cas9 against diverse group of ssDNA viruses.

plant biology↗

Toward More General Embeddings for Protein Design: Harnessing Joint Representations of Sequence and Structure

AO_SCPLOWBSTRACTC_SCPLOWProtein embeddings learned from aligned sequences have been leveraged in a wide array of tasks in protein understanding and engineering. The sequence embeddings are generated through semi-supervised training on millions of sequences with deep neural models defined with hundreds of millions of parameters, and they continue to increase in performance on target tasks with increasing complexity. We report a more data-efficient approach to encode protein information through joint training on protein sequence and structure in a semi-supervised manner. We show that the method is able to encode both types of information to form a rich embedding space which can be used for downstream prediction tasks. We show that the incorporation of rich structural information into the context under consideration boosts the performance of the model by predicting the effects of single-mutations. We attribute increases in accuracy to the value of leveraging proximity within the enriched representation to identify sequentially and spatially close residues that would be affected by the mutation, using experimentally validated or predicted structures.

biochemistry↗

An ERK5-KLF2 signalling module regulates early embryonic gene expression dynamics and stem cell rejuvenation

The ERK5 MAP kinase signalling pathway drives transcription of naive pluripotency genes in mouse Embryonic Stem Cells (mESCs). However, how ERK5 impacts on other aspects of mESC biology has not been investigated. Here, we employ quantitative proteomic profiling to identify proteins whose expression is regulated by the ERK5 pathway in mESCs. This reveals a function for ERK5 signalling in regulating dynamically expressed early embryonic 2-cell stage (2C) genes including the mESC rejuvenation factor ZSCAN4. ERK5-dependent ZSCAN4 induction in mESCs increases telomere length, a key rejuvenative process required for prolonged culture. Mechanistically, ERK5 promotes ZSCAN4 and 2C gene expression via transcription of the KLF2 pluripotency transcription factor. Surprisingly, ERK5 also directly phosphorylates KLF2 to drive ubiquitin-dependent degradation, encoding negative-feedback regulation of 2C gene expression. In summary, our data identify a regulatory module whereby ERK5 kinase and transcriptional activities bi-directionally control KLF2 levels to pattern dynamic 2C gene transcription and mESC rejuvenation.

developmental biology↗

Genotyping Test Development and Genotyping Survey of Pakistani population of Holstein Friesian imported from Different Origins for A1/A2 SNP in Beta-casein Gene

Quality of milk is determined by multiple factors including microbial load, somatic cell score, content percentages and absence of adulterants. However, in term of genetics, milk quality can be linked with beta casein. Beta casein is a milk protein having genetic variants, one of which is believed to be linked with progression of many diseases including cardiovascular diseases, metabolic diseases and nervous disorders. As beta casein has genetic basis so milk quality can be elucidated by animal genotyping. But no such genotyping facility was available country wide for this purpose. Hence objective of this study is the development of diagnostic test using ARMS-PCR technology in Pakistan and genotyping of beta casein variants in Pakistani population of Holstein Friesian cattle. A total of 166 blood samples were collected from Holstein Friesian (imported from Holland, USA and Australia) and genotyped using ARMS-PCR. Validation of ARMS-PCR was confirmed by Sanger sequencing of beta casein gene from 78 animals. ARMS-PCR based test was then compared with parallel genotyping techniques in term of cost and time effectivity. Reproducibility of this test was tested and validation of this test was done by Sanger sequencing. This test is cost effective as well as time effective in compared to other techniques. Holstein Friesian originating from different countries have different percentages of beta casein variants. This test can be used routinely for genotyping of beta casein. In future there is a need to find out prevalence of beta casein variants of indigenous cattle as well as cross bred population of Pakistan and formulation of breeding policy, keeping in view health concerns of A1 genotype of {beta}-casein.\n\nNovelty StatementIn this study, a diagnostic test is developed to genotype beta casein variants in animal. This is the first report of its kind in Pakistan as before this report either sequencing or tedious and costly procedures were in use for genotyping of beta casein variants in Pakistan. Additionally, for the first time in Pakistan Holstein Friesian imported from Holland, United States and Australia were genotyped to elucidate their genotype status for beta casein polymorphism.

genetics↗

Silencing cathepsin L expression reduces Myzus persicae protein content and the nutritional value as prey for Coccinella septempunctata

Gut-expressed aphid genes, which may be more easily inhibited by RNA interference (RNAi) constructs, are attractive targets for pest control efforts involving transgenic plants. Here we show that expression of cathepsin L, a cysteine protease that functions in aphid guts, can be reduced by expression of an RNAi construct in transgenic tobacco. The effectiveness of this approach is demonstrated by up to 80% adult mortality, reduced fecundity, and delayed nymph production of Myzus persicae (green peach aphids) when cathepsin L expression was reduced by plant-mediated RNAi. Consistent with the function of cathepsin L as a gut protease, M. persicae fed on the RNAi plants had a lower protein content in their bodies and excreted more protein in their honeydew. Larvae of Coccinella septempunctata (seven-spotted ladybugs) grew more slowly on aphids having reduced cathepsin L expression, suggesting that prey insect nutritive value, and not just direct negative effects of the RNAi construct, needs to be considered when producing transgenic plants for RNAi-mediated pest control.\n\nHighlightsO_LISilencing expression of cathepsin L by RNA interference reduces protein content of Myzus persicae (green peach aphid) bodies.\nC_LIO_LIHoneydew of aphids with cathepsin L silenced contains elevated protein.\nC_LIO_LICathepsin L is required for efficient protein uptake from phloem sap.\nC_LIO_LIAphids with cathepsin L expression silenced have increased mortality and fewer offspring.\nC_LIO_LICoccinella septempunctata (seven-spotted ladybugs) grow more slowly on aphids with expression of cathepsin L silenced.\nC_LI\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=173 SRC=\"FIGDIR/small/451211_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (26K):\norg.highwire.dtl.DTLVardef@5ee6d7org.highwire.dtl.DTLVardef@853fe8org.highwire.dtl.DTLVardef@26c2aborg.highwire.dtl.DTLVardef@b67e2b_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗