Name
Identification of host-cellular determinants interacting with rotavirus NSP5: Positive role of ACLY in rotavirus infection
Presenter
Ranjana Sharma, ICMR - National Institute for Research in Bacterial Infections
Co-Author(s)
Ranjana Sharma1, Shreya Banerjee2, Mamta Chawla-Sarkar1*
1 Division of Virology, ICMR - National Institute for Research in Bacterial Infections, Kolkata, WB, India
2Division of Virology, Immunology & Microbiology, Chobanian & Avedisian School of Medicine, Boston University
*Correspondence: chawlam70@gmail.com
Abstract Category
Combatting and Exploiting dsRNA viruses
Abstract
Background: Rotavirus (RV), a non-enveloped dsRNA virus causes acute diarrheal disease in children (≤5 yrs). While vaccines have reduced mortality, they are less effective in low middle-income countries, making it crucial to understand host factors that facilitate RV infection for drug development to combat rotaviral diarrhea. RV forms replication hubs called viroplasms (VMs) using viral proteins NSP2 and NSP5. However, the role of host proteins in RV infection and VM dynamics remains largely unexplored. Unlike prior research that examined NSP5-host protein interaction using NSP5 transfected cells or focused only on NSP5 post-translational modifications via mass spectrometry, this study analyzed NSP5-host protein interaction in MA104 cells during RV infection.
Methods: MA104 cells were either mock-infected or infected in triplicate with RV-SA11 at 6hpi, co-immunoprecipitated using anti-NSP5 antibody followed by mass spectrometry. Bioinformatics tools were then used to analyze protein-protein interaction network and functions.
Results: Over 120 host proteins found to interact with RV-NSP5, were mainly involved in metabolic and cellular processes. Since lipids are crucial for RV infection, host protein ATP-citrate lyase (ACLY), a key enzyme involved in lipid synthesis pathway was further validated and studied. ACLY interacted with NSP5 and colocalized within VMs. RV infection increases the enzymatic activity of ACLY by phosphorylating ACLY at ser455 residue. Inhibiting ACLY, resulted in reduced infection both in-vitro and in-vivo suckling mice.
Conclusion: This study sheds light on essential cellular factors like ACLY which support RV replication highlighting its potential as a promising anti-rotaviral target.
Methods: MA104 cells were either mock-infected or infected in triplicate with RV-SA11 at 6hpi, co-immunoprecipitated using anti-NSP5 antibody followed by mass spectrometry. Bioinformatics tools were then used to analyze protein-protein interaction network and functions.
Results: Over 120 host proteins found to interact with RV-NSP5, were mainly involved in metabolic and cellular processes. Since lipids are crucial for RV infection, host protein ATP-citrate lyase (ACLY), a key enzyme involved in lipid synthesis pathway was further validated and studied. ACLY interacted with NSP5 and colocalized within VMs. RV infection increases the enzymatic activity of ACLY by phosphorylating ACLY at ser455 residue. Inhibiting ACLY, resulted in reduced infection both in-vitro and in-vivo suckling mice.
Conclusion: This study sheds light on essential cellular factors like ACLY which support RV replication highlighting its potential as a promising anti-rotaviral target.