Two non-structural rotavirus proteins, NSP2 and NSP5, form viroplasm-like structures in vivo

J Gen Virol. 1999 Feb;80 ( Pt 2):333-339. doi: 10.1099/0022-1317-80-2-333.

Abstract

In rotavirus-infected cells, the non-structural proteins NSP5 and NSP2 localize in complexes called viroplasms, where replication and assembly occur. Recently, we have demonstrated direct interaction of NSP5 with NSP2, and as a consequence of that, up-regulation of NSP5 hyperphosphorylation. To investigate a possible structural role for the NSP2-NSP5 interaction, we analysed the cytoplasmic distribution of the two proteins in transfected cells by immunofluorescence using specific antibodies. Here we report that NSP2 and NSP5 can drive the formation of viroplasm-like structures (VLS) in the absence of other rotaviral proteins and rotavirus replication. Several NSP5 deletion mutants were constructed and expressed in combination with NSP2. Both the N- and C-terminal domains of NSP5 were found to be essential for VLS formation. Only one mutant, with an internal deletion of residues 81-130, was able to interact with NSP2 to form VLS. Analysis of the phosphorylation capacity of the different mutants in vivo indicated that hyperphosphorylation of NSP5 is necessary, but not sufficient, for VLS formation. Our results suggest a role for the non-structural protein NSP5 in the structure of viroplasms mediated by its interaction with NSP2.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • DNA Primers / genetics
  • Macromolecular Substances
  • Microscopy, Fluorescence
  • Phosphorylation
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Rotavirus / genetics
  • Rotavirus / metabolism*
  • Sequence Deletion
  • Viral Nonstructural Proteins / chemistry*
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism*
  • Viral Proteins / chemistry*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*
  • Virus Replication

Substances

  • DNA Primers
  • Macromolecular Substances
  • RNA-Binding Proteins
  • Viral Nonstructural Proteins
  • Viral Proteins
  • NS35 protein, rotavirus
  • gene 11 protein, Rotavirus B