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1.
J Virol ; 75(19): 9345-56, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11533198

ABSTRACT

The subcellular localization of transmissible gastroenteritis virus (TGEV) and mouse hepatitis virus (MHV) (group I and group II coronaviruses, respectively) nucleoproteins (N proteins) were examined by confocal microscopy. The proteins were shown to localize either to the cytoplasm alone or to the cytoplasm and a structure in the nucleus. This feature was confirmed to be the nucleolus by using specific antibodies to nucleolin, a major component of the nucleolus, and by confocal microscopy to image sections through a cell expressing N protein. These findings are consistent with our previous report for infectious bronchitis virus (group III coronavirus) (J. A. Hiscox et al., J. Virol. 75:506-512, 2001), indicating that nucleolar localization of the N protein is a common feature of the coronavirus family and is possibly of functional significance. Nucleolar localization signals were identified in the domain III region of the N protein from all three coronavirus groups, and this suggested that transport of N protein to the nucleus might be an active process. In addition, our results suggest that the N protein might function to disrupt cell division. Thus, we observed that approximately 30% of cells transfected with the N protein appeared to be undergoing cell division. The most likely explanation for this is that the N protein induced a cell cycle delay or arrest, most likely in the G(2)/M phase. In a fraction of transfected cells expressing coronavirus N proteins, we observed multinucleate cells and dividing cells with nucleoli (which are only present during interphase). These findings are consistent with the possible inhibition of cytokinesis in these cells.


Subject(s)
Cell Nucleolus/virology , Coronavirus/physiology , Nucleocapsid Proteins , Nucleocapsid/physiology , Animals , Cell Line , Cell Nucleolus/immunology , Coronavirus Nucleocapsid Proteins , Fluorescent Antibody Technique, Indirect , Immunity, Innate , Mice , Virus Replication/immunology
2.
J Virol ; 75(1): 506-12, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11119619

ABSTRACT

The coronavirus nucleoprotein (N) has been reported to be involved in various aspects of virus replication. We examined by confocal microscopy the subcellular localization of the avian infectious bronchitis virus N protein both in the absence and in the context of an infected cell and found that N protein localizes both to the cytoplasmic and nucleolar compartments.


Subject(s)
Cell Nucleolus/chemistry , Infectious bronchitis virus/chemistry , Nucleocapsid Proteins/analysis , Amino Acid Sequence , Animals , Chlorocebus aethiops , Cytoplasm/chemistry , Microscopy, Confocal , Molecular Sequence Data , Vero Cells
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