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Gupta, T.

Publications and source records attributed to Gupta, T..

24 records · Page 2Linked to original sources

Molecular Basis for CPC-Sgo1 Interaction: Implications for Centromere Localisation and Function of the CPC

The Chromosomal Passenger Complex (CPC; consisting of Borealin, Survivin, INCENP and Aurora B kinase) and Shugoshin 1 (Sgo1) are key regulators of chromosome bi-orientation, a process essential for error-free chromosome segregation. Their functions rely on their ability to associate with centromeres. Two histone phosphorylations, histone H3 Thr3 (H3T3ph; directly recognised by Survivin) and histone H2A Thr120 (H2AT120ph; indirectly recognised via Sgo1), together with CPCs intrinsic ability to bind nucleosome, facilitate CPC centromere recruitment. The molecular basis for CPC-Sgo1 binding and how their direct interaction influences CPC centromere localisation and function are lacking. Here, using an integrative structure-function approach, we show that the histone H3-like Sgo1 N-terminal tail interacts with Survivin acting as a hot-spot for CPC-Sgo1 assembly, while downstream Sgo1 residues, mainly with Borealin contributes for high affinity interaction. Disruption of the Sgo1 N-terminal tail-Survivin interaction abolished CPC-Sgo1 assembly in vitro and perturbed centromere localisation and function of CPC. Our findings provide evidence that CPC binding to Sgo1 and histone H3 N-terminal tail are mutually exclusive, suggesting that these interactions will likely take place in a spatially/temporally restricted manner and provide a rationale for the Sgo1-mediated kinetochore proximal centromere pool of CPC.

cell biology↗

Identification of novel pulmonary vein nodes as generators of ectopic arrhythmic foci for atrial fibrillation: An immuno-histochemical proof

IntroductionThe atrial muscle sleeve (AMS) of the pulmonary vein is the most common source of the arrhythmogenic triggers in atrial fibrillation (AF). Anatomical substrate generating these ectopic currents is still elusive. The present study was designed to study the AMS of pulmonary veins with an emphasis on the structural basis which might govern AF initiation and perpetuation. Materials and MethodsThe study was conducted on longitudinal tissue section of pulmonary vein, taken from 15 human cadaveric non-diseased hearts. Tissue was studied histologically using H&E and Gomori trichrome stain. The pacemaker channels were identified by immunohistochemistry using monoclonal HCN4 and HCN1 antibodies. ResultsThe AMS was identified in each pulmonary vein, located between the tunica adventitia and tunica media. A node like arrangement of myocytes was seen within the AMS in 30% of veins. It had a compact zone limited by a fibrous capsule and contained much smaller, paler and interconnected myocytes. Outside the capsule there was a zone of dispersed, singly placed myocytes separating the compact zone from the working myocytes of the AMS. HCN4 and HCN1 antibodies were expressed on the cell membrane of nodal myocytes, while the working myocytes demonstrated none to minimal staining. ConclusionPulmonary veins nodes are similar to the specialized cardiac conductive tissue in, histological arrangement of compact and transitional zones, cellular characteristics, and the presence of pacemaker channels. They might be the anatomical basis of ectopic arrhythmogenic foci. To our knowledge these nodes are being described for the first time in human.

pathology↗

A highly conserved binding pocket on PP2A-B56 is required for shugoshin binding and cohesion protection

The shugoshin proteins are universal protectors of centromeric cohesin during mitosis and meiosis. The binding of human Sgo1 to the PP2A-B56 phosphatase through a coiled coil (CC) region is believed to mediate cohesion protection during mitosis. Here we undertook a structure function analysis of the PP2A-B56-Sgo1 complex, revealing unanticipated aspects of complex formation and function. We establish that a highly conserved pocket of the B56 regulatory subunit is required for Sgo1 binding and cohesion protection. Consistent with this, we show that Sgo1 blocks the binding of PP2A-B56 substrates containing a canonical B56 binding motif. Surprisingly, we identify B56 and Sgo1 mutants that prevent complex formation yet support cohesion protection and normal mitotic progression. This suggests that Sgo1 and PP2A-B56 have cohesion protection activity independently of complex formation. Collectively our work provides important insight into cohesion protection during mitosis.

cell biology↗

Role of Rv3351c in trafficking Mycobacterium tuberculosis bacilli in alveolar epithelial cells and its contribution to disease

Although interactions with alveolar macrophages have been well characterized for Mycobacterium tuberculosis, the roles epithelial cells play during infection and disease development have been less studied. We have previously shown that deletion of gene rv3351c reduces M. tuberculosis replication in and necrosis of A549 human type II pneumocyte cells. In the present study, we report that rv3351c is required for lipid raft aggregation on A549 cell plasma membranes during M. tuberculosis infection. Lipid raft aggregation was also induced directly by recombinant Rv3351c protein. A{Delta} rv3351c deletion mutant was less effective than wild type M. tuberculosis at circumventing phagolysosome fusion in A549 cells as evidenced by increased co-localization with lysosomal markers LAMP-2 and cathepsin-L by the mutant bacilli. These observations indicate a role for Rv3351c in modification of the plasma membrane to facilitate trafficking and survival of M. tuberculosis bacilli through alveolar epithelial cells, and support the hypothesis that M. tuberculosis has mechanisms to target the alveolar epithelium. Preliminary data also demonstrate that like the type II pneumocyte-targeting M. tuberculosis secreted protein heparin-binding filamentous hemagglutinin (HBHA), Rv3351c is detected by the host cellular and humoral immune responses during infection, and may play an important role in mycobacterial dissemination from the lungs. Author summaryMycobacterium tuberculosis is the leading causes of death due to a single infectious agent and many facets regarding the pathogenesis of this organism remain unknown. This facultative intracellular bacterial pathogen often establishes infection through inhalation of the bacilli into the alveoli of the lungs. Interactions with alveolar macrophages have been well characterized and it had been assumed that these interactions with phagocytic cells primarily determine the fate of the disease. However, alveolar epithelial cells, such as type II pneumocytes, play important roles in disease progression of other bacterial and viral respiratory pathogens, which provided the impetus to more-closely examine pneumocyte-M. tuberculosis interactions. We describe in this study the role of the M. tuberculosis rv3351c gene product in the internalization and survival of this pathogen in human type II pneumocytes. We previously showed that a{Delta} rv3351c mutant replicates less efficiently and generates less necrosis than the parental M. tuberculosis strain in this cell type. We demonstrate herein that Rv3351c protein induces lipid raft aggregation on the membranes of alveolar epithelial cells and that M. tuberculosis {Delta}rv3351c traffics through LAMP-2-labeled endosomes 30% more frequently than the parent strain. This trafficking toward phagolysosomes may underlie the reduced replication and cytotoxicity of the mutant. The role of Rv3351c in trafficking and survival of M. tuberculosis bacilli through epithelial cells ultimately resulting in dissemination from the lungs may begin with modifications to the plasma membrane prior to attachment. Such a mechanism of activity suggests Rv3351c as a potential vaccine target to train the host immune system to bind and eliminate the protein before it modulates the alveolar epithelium.

microbiology↗

Aggression and discrimination among closely versus distantly related species of Drosophila

Fighting between different species is widespread in the animal kingdom, yet this phenomenon has been relatively understudied in the field of aggression research. Particularly lacking are studies that test the effect of genetic distance, or relatedness, on aggressive behavior between species. Here we characterized male-male aggression within and between species of fruit flies across the Drosophila phylogeny. We show that male Drosophila discriminate between conspecifics and heterospecifics and show a bias for the target of aggression that depends on the genetic relatedness of opponent males. Specifically, males of closely related species treated conspecifics and heterospecifics equally, whereas males of distantly related species were overwhelmingly aggressive toward conspecifics. To our knowledge, this is the first study to quantify aggression between Drosophila species and to establish a behavioral bias for aggression against conspecifics versus heterospecifics. Our results suggest that future study of heterospecific aggression behavior in Drosophila is warranted to investigate the degree to which these trends in aggression among species extend to broader behavioral, ecological, and evolutionary contexts.

animal behavior and cognition↗

Disparate patterns of thermal adaptation between life stages in temperate vs. tropical populations of Drosophila melanogaster

Many terrestrial ectothermic species exhibit limited variation in upper thermal tolerance across latitude. However, these trends may not signify limited adaptive capacity to increase thermal tolerance in the face of climate change. Instead, thermal tolerance may be similar among populations because behavioral thermoregulation by mobile organisms or life stages may buffer natural selection for thermal tolerance. We compared thermal tolerance of adults and embryos among natural populations of Drosophila melanogaster from a broad range of thermal habitats around the globe to assess natural variation of thermal tolerance in mobile vs. immobile life stages. We found no variation among populations in adult thermal tolerance, but embryonic thermal tolerance was higher in tropical strains than in temperate strains. Average maximum temperature of the warmest month of the year predicted embryonic thermal tolerance in tropical but not temperate sites. We further report that embryos live closer to their upper thermal limits than adultso--i.e., thermal safety margins are smaller for embryos than adults. F1 hybrid embryos from crosses between temperate and tropical populations had thermal tolerance that matched that of tropical embryos, suggesting dominance of heat-tolerant alleles. Together our findings suggest that thermal selection has led to divergence in embryonic thermal tolerance but that selection for divergent thermal tolerance may be limited in adults. Further, our results suggest that thermal traits should be measured across life stages in order to better predict adaptive limits.\n\nImpact SummaryClimate change may threaten the extinction of many ectothermic species, unless populations can evolutionarily adapt to rising temperatures. Natural selection should favor individuals with higher heat tolerances in hotter environments. But recent studies have found that individuals from hot and cold places often have similar heat tolerances. This pattern may indicate that the evolution of heat tolerance is constrained. If this were true, then it would have dire consequences for species persistence under novel thermal conditions.\n\nAn alternative explanation for lack of variation in heat tolerance is that mobile organisms dont need higher heat tolerances to survive in hotter places. The majority of studies have focused on heat tolerance of the adult life stage. Yet, adults in many species are mobile organisms that can avoid extreme heat by seeking shelter in cooler microhabitats (e.g., shaded locations). In contrast, immobile life stages (e.g., insect eggs) cannot behaviorally avoid extreme heat. Thus, mobile and immobile life stages may face different thermal selection pressures that lead to disparate patterns of thermal adaptation across life stages.\n\nHere, we compared heat tolerances of fruit fly adults and eggs (Drosophila melanogaster) from populations in temperate North America and tropical locations around the globe. Consistent with previous studies, we found no differences among populations in adult heat tolerance. However, eggs from tropical flies were consistently more heat tolerant than eggs from North American flies. Further, eggs had lower heat tolerance than adults. Consequently, fly eggs in the hotter tropics may experience heat death more frequently than adult flies later in life. This may explain why patterns of divergence in heat tolerance were decoupled across life stages. These patterns indicate that thermal adaptation may be life-stage-specific and suggest that future work should characterize thermal traits across life stages to better understand the evolution of thermal limits.

evolutionary biology↗