Identification of cellular calcium binding protein calmodulin as a regulator of rotavirus A infection during comparative proteomic study.

Chattopadhyay S, Basak T, Nayak MK, Bhardwaj G, Mukherjee A, Bhowmick R, Sengupta S, Chakrabarti O, Chatterjee NS, Chawla-Sarkar M
PLoS One. 2013 8 (2): e56655

PMID: 23437200 · PMCID: PMC3577757 · DOI:10.1371/journal.pone.0056655

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.

MeSH Terms (16)

Animals Antigens, Viral Calcium Calmodulin Capsid Proteins Dysentery Egtazic Acid HT29 Cells Humans Mice Mice, Inbred BALB C Protein Binding Rotavirus Rotavirus Infections Two-Dimensional Difference Gel Electrophoresis Virus Replication

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