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Tomar, R. S.

Publications and source records attributed to Tomar, R. S..

2 recordsLinked to original sources

A novel link between Cell Wall Integrity pathway and Flocculation, regulated by yeast Sen1 dependent interplay between Rlm1 and Tup1

The budding yeast, Saccharomyces cerevisiae is one of the most studied organisms used for the synthesis of products, to explore the human diseases and eukaryotic gene expression mechanisms. The yeast cells with flocculation property are in high demand for industrial applications. However, the pathogenic yeast becomes drug-resistant due to flocculation/biofilm phenotype. The flocculation property of yeast depends on the expression of specific FLO genes. Genetic and epigenetic factors have been suggested to induce the expression of FLOs and flocculation, an evolutionarily conserved process. The present study was undertaken to identify a molecular link between stress caused by genetic and epigenetic factors and expression of FLOs. We utilized flocculating yeast strains to study the regulation of FLO genes and flocculation phenotype. We found rough surface morphology and constitutive activation of Slt2 in flocculating cells. The external cell wall stress factors as well as specific mutations in Sen1 and histone proteins strongly correlated with the induction of FLO genes whereas deletion of SLT2/RLM1, suppressed the expression and flocculation phenotype. We detected constitutive binding of Rlm1 and eviction of Tup1 from the promoters of FLO1 and FLO5 genes in flocculating cells. Thus we provide evidence for the CWI pathway dependent flocculation of yeast, regulated by Sen1 mediated interplay between Tup1 and Rlm1.

molecular biology

Cantharidin alters GPI-anchored protein sorting by targeting Cdc1 mediated remodeling in Endoplasmic Reticulum

Cantharidin (CTD) is a potent anticancer small molecule produced by several species of blister beetle. It has been a traditional medicine for the treatment of warts and tumors for many decades. CTD suppresses the tumor growth by inducing apoptosis, cell cycle arrest, and DNA damage. It is a known inhibitor of PP2A and PP1. In this study, we identified new molecular targets of CTD using Saccharomyces cerevisiae as a model organism which expresses a Cantharidin Resistance Gene (CRG1). CRG1 encodes a SAM-dependent methyltransferase that inactivates CTD by methylation. CTD alters lipid homeostasis, cell wall integrity, endocytosis, adhesion, and invasion in yeast cells. We found that CTD specifically affects the phosphatidylethanolamine (PE) associated functions which can be rescued by supplementation of ethanolamine (ETA) in the growth media. CTD also perturbed ER homeostasis and cell wall integrity by altering the GPI-anchored protein sorting. The CTD dependent genetic interaction profile of CRG1 revealed that Cdc1 activity in GPI-anchor remodeling is the key target of CTD, which we found to be independent of PP2A and PP1. Furthermore, our experiments with human cells suggest that CTD functions through a conserved mechanism in higher eukaryotes as well. Altogether, we conclude that CTD induces cytotoxicity by targeting Cdc1 activity in GPI-anchor remodeling in the endoplasmic reticulum (ER).

cell biology