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Journal of Virology, May 2002, p. 5233-5250, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.5233-5250.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Interaction of the Coronavirus Nucleoprotein with Nucleolar Antigens and the Host Cell

Hongying Chen,1 Torsten Wurm,1 Paul Britton,2 Gavin Brooks,3 and Julian A. Hiscox1*

Virology,1 Cell Cycle Groups, School of Animal and Microbial Sciences, The University of Reading, Reading,3 Division of Molecular Biology, Institute for Animal Health, Compton, United Kingdom2

Received 10 December 2001/ Accepted 6 February 2002

Coronavirus nucleoproteins (N proteins) localize to the cytoplasm and the nucleolus, a subnuclear structure, in both virus-infected primary cells and in cells transfected with plasmids that express N protein. The nucleolus is the site of ribosome biogenesis and sequesters cell cycle regulatory complexes. Two of the major components of the nucleolus are fibrillarin and nucleolin. These proteins are involved in nucleolar assembly and ribosome biogenesis and act as chaperones for the import of proteins into the nucleolus. We have found that fibrillarin is reorganized in primary cells infected with the avian coronavirus infectious bronchitis virus (IBV) and in continuous cell lines that express either IBV or mouse hepatitis virus N protein. Both N protein and a fibrillarin-green fluorescent protein fusion protein colocalized to the perinuclear region and the nucleolus. Pull-down assays demonstrated that IBV N protein interacted with nucleolin and therefore provided a possible explanation as to how coronavirus N proteins localize to the nucleolus. Nucleoli, and proteins that localize to the nucleolus, have been implicated in cell growth-cell cycle regulation. Comparison of cells expressing IBV N protein with controls indicated that cells expressing N protein had delayed cellular growth. This result could not to be attributed to apoptosis. Morphological analysis of these cells indicated that cytokinesis was disrupted, an observation subsequently found in primary cells infected with IBV. Coronaviruses might therefore delay the cell cycle in interphase, where maximum translation of viral mRNAs can occur.

* Corresponding author. Mailing address: School of Animal and Microbial Sciences, University of Reading, Whiteknights, P.O. Box 228, Reading RG6 6AJ, United Kingdom. Phone: (44)118-931-8893. Fax: (44)118-931-0180. E-mail: j.a.hiscox{at}reading.ac.uk.

Journal of Virology, May 2002, p. 5233-5250, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.5233-5250.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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