Biology subjects

Guha, A.

Publications and source records attributed to Guha, A..

4 recordsLinked to original sources

bioRxiv2021

The Fragile X Mental Retardation Protein protects the lung from xenobiotic stress by facilitating the Integrated Stress Response

Stress response pathways protect the lung from the damaging effects of environmental toxicants. Here we investigate the role of the Fragile X Mental Retardation Protein (FMRP), a multifunctional protein implicated in stress responses, in the lung. We report that FMRP is expressed in murine and human lungs, in the airways and more broadly. Analysis of airway stress responses in mice and in a murine cell line ex vivo, using the well-established Naphthalene (Nap) injury model, reveals that FMRP-deficient cells exhibit increased expression of markers of oxidative and genotoxic stress and increased cell death. We find that FMRP-deficient cells fail to actuate the Integrated Stress Response Pathway (ISR) and upregulate the transcription factor ATF4. Knockdown of ATF4 expression phenocopies the loss of FMRP. We extend our analysis of the role of FMRP to human bronchial BEAS-2B cells, using a 9, 10-Phenanthrenequinone air pollutant model, to find that FMRP-deficient BEAS-2B also fail to actuate the ISR and exhibit greater susceptibility. Taken together, our data suggest that FMRP has a conserved role in protecting the airways by facilitating the ISR.

cell biology↗

bioRxiv2020

Niclosamide inhibits SARS-CoV2 entry by blocking internalization through pH-dependent CLIC/GEEC endocytic pathway

Many viruses utilize the host endo-lysosomal network to infect cells. Tracing the endocytic itinerary of SARS-CoV2 can provide insights into viral trafficking and aid in designing new therapeutic targets. Here, we demonstrate that the receptor binding domain (RBD) of SARS-CoV2 is internalized via the clathrin and dynamin-independent, pH-dependent CLIC/GEEC (CG) endocytic pathway. Endosomal acidification inhibitors like BafilomycinA1 and NH4Cl, which inhibit the CG pathway, strongly block the uptake of RBD. Using transduction assays with SARS-CoV2 Spike-pseudovirus, we confirmed that these acidification inhibitors also impede viral infection. By contrast, Chloroquine neither affects RBD uptake nor extensively alters the endosomal pH, yet attenuates Spike-pseudovirus entry, indicating a pH-independent mechanism of intervention. We screened a subset of FDA-approved acidification inhibitors and found Niclosamide to be a potential SARS-CoV2 entry inhibitor. Niclosamide, thus, could provide broader applicability in subverting infection of similar category viruses entering host cells via this pH-dependent endocytic pathway.

cell biology↗

bioRxiv2020

Response of whole plant water use to limiting light and water conditions are independent of each other in seedlings of seasonally dry tropical forests

How co-occurring species vary in the utilization of a shared and limited supply of water, especially in the context of other limiting resources like light, is essential for understanding processes that facilitate species coexistence and community assembly. For seedlings in a seasonally dry tropical forest that experience large heterogeny in light and water conditions, how water use, leaf physiology, and subsequently plant growth, is affected by limited water and light availability is still not well understood. In a controlled common garden experiment with four co-existing and commonly occurring dry tropical forest species, we examined how whole plant water uptake, responds to limiting water and light conditions and whether these responses are reflected in leaf physiology, and translated to growth. Water use varied dramatically in seedlings of the four species with a five-fold difference in well-watered plants grown in full sunlight. Species varied in their response to shade, but did not differ in responses to the low water treatment, possibly resulting from the strong selective force imposed by the very low water availability and the long dry period characteristic of these seasonally dry forests. Interestingly, species responses in water use, physiology, and growth in limiting water conditions were independent of light. Thus, species response to both these limiting conditions may evolve independently of each other. Responses in water use were largely congruent with responses in leaf physiology and growth. However, while magnitude of changes in leaf physiology were largely driven by light conditions, changes in whole plant water use and growth were influenced to a greater degree by the water treatment. This highlights the need to measure whole plant water use to better understand plant growth responses in these seasonally dry tropical forests.

ecology↗

bioRxiv2020

Multiple Wnts act synergistically to induce Chk1/Grapes expression and mediate G2 arrest in Drosophila tracheoblasts

Larval tracheae of Drosophila harbor progenitors of the adult tracheal system (tracheoblasts). Thoracic tracheoblasts are arrested in the G2 phase of the cell cycle in an ATR (mei-41)-Checkpoint Kinase1 (grapes, Chk1) dependent manner prior to mitotic re-entry. Here we investigate developmental regulation of Chk1 activation. We report that Wnt signaling is high in tracheoblasts and is necessary for high levels of activated (phosphorylated) Chk1. We find that canonical Wnt signaling facilitates this by transcriptional upregulation of Chk1 expression in cells that have ATR kinase activity. Wnt signalling is dependent on four Wnts (Wg, Wnt5, 6,10) that are expressed at high levels in arrested tracheoblasts and downregulated at mitotic re-entry. Interestingly, none of the Wnts are dispensable and act synergistically to induce Chk1. Finally, we show that downregulation of Wnt signalling and Chk1 expression leads to mitotic re-entry and the concomitant upregulation of Dpp signalling, driving tracheoblast proliferation.

developmental biology↗