title: Comprehensive analysis of temporal alterations in cellular proteome of Bacillus subtilis under curcumin treatment.
abstract: Curcumin is a natural dietary compound with antimicrobial activity against various gram positive and negative bacteria. This study aims to investigate the proteome level alterations in Bacillus subtilis due to curcumin treatment and identification of its molecular/cellular targets to understand the mechanism of action. We have performed a comprehensive proteomic analysis of B. subtilis AH75 strain at different time intervals of curcumin treatment (20, 60 and 120 min after the drug exposure, three replicates) to compare the protein expression profiles using two complementary quantitative proteomic techniques, 2D-DIGE and iTRAQ. To the best of our knowledge, this is the first comprehensive longitudinal investigation describing the effect of curcumin treatment on B. subtilis proteome. The proteomics analysis revealed several interesting targets such UDP-N-acetylglucosamine 1-carboxyvinyltransferase 1, putative septation protein SpoVG and ATP-dependent Clp protease proteolytic subunit. Further, in silico pathway analysis using DAVID and KOBAS has revealed modulation of pathways related to the fatty acid metabolism and cell wall synthesis, which are crucial for cell viability. Our findings revealed that curcumin treatment lead to inhibition of the cell wall and fatty acid synthesis in addition to differential expression of many crucial proteins involved in modulation of bacterial metabolism. Findings obtained from proteomics analysis were further validated using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) assay for respiratory activity, resazurin assay for metabolic activity and membrane integrity assay by potassium and inorganic phosphate leakage measurement. The gene expression analysis of selected cell wall biosynthesis enzymes has strengthened the proteomics findings and indicated the major effect of curcumin on cell division.
authors: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors: Panga Jaipal
authors: PJ
authors: Reddy
authors: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors: Sneha
authors: S
authors: Sinha
authors: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors: Sandipan
authors: S
authors: Ray
authors: Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, India; Manipal University, Madhav Nagar,Manipal, India.
authors: Gajanan J
authors: GJ
authors: Sathe
authors: Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, India; Manipal University, Madhav Nagar,Manipal, India.
authors: Aditi
authors: A
authors: Chatterjee
authors: Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, India.
authors: T S Keshava
authors: TS
authors: Prasad
authors: Proteomics Laboratory, National Centre for Cell Science, Ganeshkhind, Pune, Maharashtra, India.
authors: Snigdha
authors: S
authors: Dhali
authors: Proteomics Laboratory, National Centre for Cell Science, Ganeshkhind, Pune, Maharashtra, India.
authors: Rapole
authors: R
authors: Srikanth
authors: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors: Dulal
authors: D
authors: Panda
authors: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors: Sanjeeva
authors: S
authors: Srivastava
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D001426:Bacterial Proteins / Q000235:genetics; D002473:Cell Wall / Q000378:metabolism; D003198:Computer Simulation; D003474:Curcumin / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005227:Fatty Acids / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D008954:Models, Biological; D010457:Peptidoglycan / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D020543:Proteome / Q000187:drug effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D005227:Fatty Acids; D010457:Peptidoglycan; D010710:Phosphates; D020543:Proteome; D003474:Curcumin; D011188:Potassium