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Rotavirus NSP5 phosphorylation is up-regulated by interaction with NSP2
I Afrikanova1, E Fabbretti, M C Miozzo
1International Centre for Genetic Engineering and Biotechnology, Padriciano, Trieste, Italy.
The Journal of General Virology
|November 20, 1998
Summary
Rotavirus non-structural protein 5 (NSP5) interacts with NSP2, influencing its phosphorylation. Deletion mutants reveal specific domains regulating NSP5 phosphorylation and NSP2 interaction, crucial for viral replication.
Area of Science:
- Virology
- Molecular Biology
- Protein Biochemistry
Background:
- Rotavirus non-structural protein 5 (NSP5) exists in multiple isoforms with varying phosphorylation levels.
- NSP5 co-localizes with NSP2 in viroplasms, sites of rotavirus replication.
- Autophosphorylation is a key mechanism regulating NSP5 phosphorylation.
Purpose of the Study:
- To investigate the interaction between rotavirus NSP5 and NSP2.
- To elucidate the role of specific NSP5 domains in phosphorylation and NSP2 interaction.
- To map NSP5 autophosphorylation activity.
Main Methods:
- Chemical cross-linking in living cells.
- Co-immunoprecipitation assays.
- Transient transfection assays with NSP5 deletion mutants.
Main Results:
- NSP5 was cross-linked with viral polymerase VP1 and NSP2 in infected cells.
- NSP5 and NSP2 interaction was confirmed by co-immunoprecipitation.
- Co-expression of NSP2 enhanced NSP5 phosphorylation in vivo.
- C-terminal deletion abolished NSP5 phosphorylation, while N-terminal deletion led to hyperphosphorylation and NSP2 insensitivity.
Conclusions:
- NSP2 positively regulates NSP5 phosphorylation.
- The N-terminal domain of NSP5 plays a critical regulatory role in phosphorylation and NSP2 interaction.
- Specific domains of NSP5 are involved in autophosphorylation and interaction with NSP2, impacting rotavirus replication.