Biology subjects

Kamat, A.

Publications and source records attributed to Kamat, A..

7 recordsLinked to original sources

bioRxiv2025

RNA triggers chronic stress during neuronal aging

Neurodegenerative diseases are linked with dysregulation of the integrated stress response (ISR), which coordinates cellular homeostasis during and after stress events. Cellular stress can arise from several sources, but there is significant disagreement about which stress might contribute to aging and neurodegeneration. Here, we leverage directed transdifferentiation of human fibroblasts into aged neurons to determine the source of ISR activation. We demonstrate that increased accumulation of cytoplasmic double-stranded RNA (dsRNA) activates the eIF2 kinase PKR, which in turn triggers the ISR in aged neurons and leads to sequestration of dsRNA in stress granules. Aged neurons accumulate endogenous mitochondria-derived dsRNA that directly binds to PKR. This mitochondrial dsRNA leaks through damaged mitochondrial membranes and forms cytoplasmic foci in aged neurons. Finally, we demonstrate that PKR inhibition leads to the cessation of stress, resumption of cellular translation, and restoration of RNA-binding protein expression. Together, our results identify a source of RNA stress that destabilizes aged neurons and may contribute to neurodegeneration.

cell biology↗

bioRxiv2025

Variability in intrinsic promoter strength underlies the temporal hierarchy of the Caulobacter SOS response induction

Bacteria encode for gene regulatory networks crucial for sensing and repairing DNA damage. Upon exposure to genotoxic stress, these transcriptional networks are induced in a temporally structured manner. A case in point is of the highly conserved SOS response thwat is regulated by the LexA repressor. Studies have proposed that affinity of LexA towards promoters of SOS response genes is the primary determinant of its expression dynamics. Here, we describe an additional level of regulation beyond LexA properties that modulates the SOS response gene expression pattern. Using transcriptomic analyses, we reveal a distinct temporal hierarchy in the induction of SOS-regulated genes in Caulobacter crescentus. We observe that LexA properties are insufficient in predicting the temporal hierarchy of these genes. Instead, we find that intrinsic promoter strength underlies the order of gene activation, with differential sigma factor association as a key factor in modulating gene expression timing. Our findings highlight a novel regulatory layer in SOS dynamics and underscore the importance of promoter properties in shaping bacterial stress responses.

microbiology↗

bioRxiv2023

Widespread prevalence of a post-translational modification in activation of an essential bacterial DNA damage response

DNA methylation plays central roles in diverse cellular processes, ranging from error-correction during replication to regulation of bacterial defense mechanisms. Nevertheless, certain aberrant methylation modifications can have lethal consequences. The mechanisms by which bacteria detect and respond to such damage remain incompletely understood. Here, we discover a highly conserved but previously uncharacterized transcription factor (Cada2), which orchestrates a methylation-dependent adaptive response in Caulobacter. This response operates independently of the SOS response, governs the expression of genes crucial for direct repair, and is essential for surviving methylation-induced damage. Our molecular investigation of Cada2 reveals a cysteine methylation-dependent post-translational modification and mode of action distinct from its E. coli counterpart, a trait conserved across all bacteria harboring a Cada2-like homolog instead. Extending across the bacterial kingdom, our findings support the notion of divergence and co-evolution of adaptive response transcription factors and their corresponding sequence-specific DNA motifs. Despite this diversity, the ubiquitous prevalence of adaptive response regulators underscores the significance of a transcriptional switch, mediated by methylation post-translational modification, in driving a specific and essential bacterial DNA damage response.

microbiology↗

bioRxiv2022

Latch Verified Bulk-RNA Seq toolkit: a cloud-based suite of workflows for bulk RNA-seq quality control, analysis, and functional enrichment

BackgroundAnalysis of high-throughput bulk RNA-sequencing (RNA-seq) data reveals changes in gene expression between diverse conditions. Many tools have emerged to quality control RNA-seq reads, quantify expression levels, conduct functional enrichment among differentially expressed genes, or identify differential RNA splicing. However, unified toolkits for conducting these analyses are lacking. Moreover, existing software does not use cloud-based platforms that provide the necessary storage and computational resources to process RNA-seq data or intuitive graphical interfaces for easy use by experimental and computational scientists. ResultsTo address these challenges, we introduce the Latch Verified Bulk RNA-Seq (LVBRS) toolkit, a flexible suite of programs packaged into a single workflow coupled with a graphical user interface for conducting quality control, transcript quantification, differential splicing, differential expression analysis, and functional enrichment analyses. For functional enrichment, the LVBRS toolkit supports three databases--Gene Ontology, KEGG Pathway, and Molecular Signatures database--capturing diverse functional information. We demonstrate the utility of the LVBRS toolkit by reanalyzing a publicly available dataset examining the impact of severe and mild models of hypoxia--induced by Cobalt (II) Chloride (CoCl2) and oxyquinoline treatment, respectively--on a human colon adenocarcinoma cell line. Our analyses reveal CoCl2 treatment results in more differentially expressed genes, recapitulating previously reported results that CoCl2 models more severe hypoxia. Moreover, including alternative splicing and functional enrichment analysis using a greater breadth of functional databases revealed additional biological insights--such as greater alternative splicing in the CoCl2 condition and differentially expressed DNA repair pathways. These results demonstrate the LVBRS toolkits efficacy in facilitating biological insights from bulk RNA-seq data. ConclusionsThe LVBRS toolkit offers a robust unified framework for processing and analyzing Bulk RNA-Seq experiments. The easy-to-use graphical user interface will enable diverse scientists to conduct high-throughput bulk RNA-Seq analysis efficiently. Our aim is that the LVBRS toolkit will help streamline bulk RNA-seq workflows and facilitate deriving biologically meaningful insights from bulk RNA-seq data. The source code is freely available under the MIT license and hosted on the LatchBio Console (https://console.latch.bio/se/bulk-rnaseq), complete with documentation (https://latch.wiki/bulk-rna-seq-end-to-end).

bioinformatics↗

bioRxiv2021

Efference information flow during skill acquisition mediates its interaction with medical simulation technology

Despite substantial progress towards establishing virtual reality (VR) simulators as a replacement for physical ones for skill training, its effect on the brain network during skill acquisition has not been well addressed. In this study, we employed portable optical neuroimaging technology and Granger causality approach to uncover the impact of the two medical simulation technologies on the directed functional brain network of the subjects with two different skill levels. The mobile brain-behavior relantionship was evaluated using functional near-infrared spectroscopy (fNIRS) while right-handed subjects performed well-established fundamentals of laparoscopic surgery (FLS) pattern cutting task. A multiple regression path analysis found that the cognitive-action information flow from the right prefrontal cortex to the supplementary motor area statistically significantly predicted the FLS task performance. Here, the skill level (expert vs novice) affected the cognitive-action information flow from the right prefrontal cortex and the efference copy information flow from the left primary motor cortex via supplementary motor area as hub to the cognitive-perception at the left prefrontal cortex, i.e., the action-preception link. The simulation technology (physical vs VR simulator) affected solely the cognitive-action information flow from the right prefrontal cortex to the left primary motor cortex; however, the interaction between the medical simulation technology) and the skill level affected the efference information flow from the left primary motor cortex to the right prefrontal cortex and from the supplementary motor area to the left prefrontal cortex. These discriminative findings are crucial since our VR simulator had face and construct validity. Therefore, our study highlighted the importance of efference information flow within the framework of the perception-action cycle when comparing medical simulation technology for visuomotor skill acquisition.

neuroscience↗

bioRxiv2021

Interhemispheric functional connectivity in the primary motor cortex distinguishes between training on a physical and a virtual surgical simulator

Functional brain connectivity using functional near-infrared spectroscopy (fNIRS) during a pattern cutting (PC) task was investigated in physical and virtual simulators. 14 right-handed novice medical students were recruited and divided into separate cohorts for physical (N=8) and virtual (N=6) PC training. Functional brain connectivity measured were based on wavelet coherence (WCOH) from task-related oxygenated hemoglobin (HBO2) changes from baseline at left and right prefrontal cortex (LPFC, RPFC), left and right primary motor cortex (LPMC, RPMC), and supplementary motor area (SMA). HBO2 changes within the neurovascular frequency band (0.01-0.07Hz) from long-separation channels were used to compute average inter-regional WCOH metrics during the PC task. The coefficient of variation (CoV) of WCOH metrics and PC performance metrics were compared. WCOH metrics from short-separation fNIRS time-series were separately compared. Partial eta squared effect size (Bonferroni correction) between the physical versus virtual simulator cohorts was found to be highest for LPMC-RPMC connectivity. Also, the percent change in magnitude-squared WCOH metric was statistically (p<0.05) different for LPMC-RPMC connectivity between the physical and the virtual simulator cohorts. Percent change in WCOH metrics from extracerebral sources was not different at the 5% significance level. Also, higher CoV for both LPMC-RPMC magnitude-squared WCOH metric and PC performance metrics were found in physical than a virtual simulator. We conclude that interhemispheric connectivity of the primary motor cortex is the distinguishing functional brain connectivity feature between the physical versus the virtual simulator cohorts. Brain-behavior relationship based on CoV between the LPMC-RPMC magnitude-squared WCOH metric and the FLS PC performance metric provided novel insights into the neuroergonomics of the physical and virtual simulators that is crucial for validating Virtual Reality technology.

neuroscience↗

bioRxiv2018

Dysregulation of EMT Drives the Progression to Clinically Aggressive Sarcomatoid Bladder Cancer

The sarcomatoid variant of urothelial bladder cancer (SARC) displays a high propensity for distant metastasis and is associated with short survival. We report a comprehensive genomic analysis of 28 cases of SARCs and 84 cases of conventional urothelial carcinomas (UCs), with the TCGA cohort of 408 muscle-invasive bladder cancers serving as the reference. SARCs showed a distinct mutational landscape with enrichment of TP53, RB1, and PIK3CA mutations. They were related to the basal molecular subtype of conventional UCs and could be divided into epithelial/basal and more clinically aggressive mesenchymal subsets based on TP63 and its target genes expression levels. Other analyses revealed that SARCs are driven by downregulation of homotypic adherence genes and dysregulation of cell cycle and EMT networks, and nearly half exhibited a heavily infiltrated immune phenotype. Our observations have important implications for prognostication and the development of more effective therapies for this highly lethal variant of bladder cancer.

genomics↗