title: Identification of cellular calcium binding protein calmodulin as a regulator of rotavirus A infection during comparative proteomic study.
abstract: Rotavirus (RV) being the major diarrhoegenic virus causes around 527000 children death (<5 years age) worldwide. In cellular environment, viruses constantly adapt and modulate to survive and replicate while the host cell also responds to combat the situation and this results in the differential regulation of cellular proteins. To identify the virus induced differential expression of proteins, 2D-DIGE (Two-dimensional Difference Gel Electrophoresis) based proteomics was used. For this, HT-29 cells were infected with RV strain SA11 for 0 hours, 3 hours and 9 hours post infection (hpi), differentially expressed spots were excised from the gel and identified using MALDI-TOF/TOF mass spectrometry. 2D-DIGE based proteomics study identified 32 differentially modulated proteins, of which 22 were unique. Some of these were validated in HT-29 cell line and in BALB/c mice model. One of the modulated cellular proteins, calmodulin (CaM) was found to directly interact with RV protein VP6 in the presence of Ca(2+). Ca(2+)-CaM/VP6 interaction positively regulates RV propagation since both CaM inhibitor (W-7) and Ca(2+) chelator (BAPTA-AM) resulted in decreased viral titers. This study not only identifies differentially modulated cellular proteins upon infection with rotavirus in 2D-DIGE but also confirmed positive engagement of cellular Ca(2+)/CaM during viral pathogenesis.
authors: Division of Virology, National Institute of Cholera and Enteric Diseases, Kolkata, West Bengal, India.
authors: Shiladitya
authors: S
authors: Chattopadhyay
authors: Trayambak
authors: T
authors: Basak
authors: Mukti Kant
authors: MK
authors: Nayak
authors: Gourav
authors: G
authors: Bhardwaj
authors: Anupam
authors: A
authors: Mukherjee
authors: Rahul
authors: R
authors: Bhowmick
authors: Shantanu
authors: S
authors: Sengupta
authors: Oishee
authors: O
authors: Chakrabarti
authors: Nabendu S
authors: NS
authors: Chatterjee
authors: Mamta
authors: M
authors: Chawla-Sarkar
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry / Q000378:metabolism*; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D004403:Dysentery / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D019073:HT29 Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D012401:Rotavirus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D012400:Rotavirus Infections / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D057825:Two-Dimensional Difference Gel Electrophoresis; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D002147:Calmodulin; D036022:Capsid Proteins; C061332:VP6 protein, Rotavirus; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D002118:Calcium