Aims
The aim of the study was to investigate the proteomic changes and antioxidant enzyme activity in chromium‐resistant Bacillus cereus VITSH1 in response to heavy metal toxicity.
Methods and Results
The variation in protein expression and antioxidant enzyme activity in the presence and absence of metal was studied in B. cereus VITSH1. The differentially expressed proteins in chromium‐treated conditions were determined by SDS PAGE. The proteins involved in metal binding, protein biosynthesis, protein folding, energy metabolism and motility were identified by mass spectrometry coupled with bioinformatics search tools. The in gel assays for antioxidant enzymes indicated a change in their activity under metal stress conditions.
Conclusions
The findings of this study suggest that the organism combats metal stress probably by restricting the entry of metal inside the cell. The role of the differentially expressed proteins clearly indicates that the first line of defence is to avoid the entry of metal into the cell either by possessing a modified outer membrane or by moving away from the toxicant. Further work on the identification of other proteins playing a role in resistance would help in integrating the available knowledge on the resistance mechanisms the organisms employ to combat toxicity.
Significance and Impact of the Study
The proteomic changes in the metal‐exposed bacteria would give an insight into the proteins involved in metal resistance mechanisms and thereby aid in the development of biosensor‐based technology for heavy metal detection.
Bacteria survive metal stress by several mechanisms and metal binding is one such mechanism which has been screened in the present study to investigate the survival strategies of metal resistant bacteria. The production of siderophores, a metal chelating agent, was detected by chrome azurol S agar assay. The changes in cell wall studied by analysing the peptidoglycan and teichoic acid content indicated an increase in the cell wall content. Evaluation of morphological and physiological alterations like cell size, granularity analysed by SEM and flow cytometry analysis revealed an increase in cell size and granularity respectively. The transformation of phosphates monitored by 31P NMR analysis indicated the presence of inorganic phosphate. Based on the cell wall changes and the 31P NMR analysis, the surface charge of the organism was studied by zeta potential which displayed a difference at pH7.
Phospholipase D (PLD) isoenzymes participate in a variety of cellular functions that are mostly attributed to phosphatidic acid (PA) synthesis. Dysregulation of PLD regulates tumor progression and metastasis, yet little is known about the underlying mechanism. We previously reported on the expression and clinical role of the PLD isoenzymes PLD1 and PLD2 in tubo-ovarian high-grade serous carcinoma (HGSC). In the present study, we investigated the biological function of PLD1 and PLD2 using the OVCAR-3 and OVCAR-8 HGSC cell lines. KO cell lines for both PLDs were generated using CRISPR/CAS9 technology and assayed for exosome secretion, spheroid formation, migration, invasion and expression of molecules involved in epithelial-mesenchymal transition (EMT) and intracellular signaling. Significant differences between PLD1 and PLD2 KO cells and controls were observed for all the above parameters, supporting an important role for PLD in regulating migration, invasion, metastasis and EMT.
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