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Journal of Virology, May 2000, p. 4807-4815, Vol. 74, No. 10
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

H-1 Parvovirus-Associated Replication Bodies: a Distinct Virus-Induced Nuclear Structure

Celina Cziepluch,1,* Stefan Lampel,2 Annabel Grewenig,1 Christine Grund,3 Peter Lichter,2 and Jean Rommelaere1

Applied Tumor Virology Unit, F0100 and Institut National de la Santé et de la Recherche Médicale U 375,1 Organization of Complex Genomes Unit,2 and Division of Cell Biology-A0100,3 Deutsches Krebsforschungszentrum, D-69120 Heidelberg, Germany

Received 13 December 1999/Accepted 17 February 2000

We have identified a nuclear structure that is induced after infection with the autonomous parvovirus H-1. Using fluorescence microscopy, we observed that the major nonstructural protein (NS1) of H-1 virus which is essential for viral DNA amplification colocalized with virus-specific DNA sequences and sites of ongoing viral DNA replication in distinct nuclear bodies which we designated H-1 parvovirus-associated replication bodies (H-1 PAR-bodies). In addition, two cellular proteins were shown to accumulate in H1 PAR-bodies: (i) the proliferating cell nuclear antigen (PCNA) which is essential for chromosomal and parvoviral replication and (ii) the NS1-interacting small glutamine-rich TPR-containing protein (SGT), suggesting a role for the latter in parvoviral replication and/or gene expression. Since many DNA viruses target preexisting nuclear structures, known as PML-bodies, for viral replication and gene expression, we have determined the localization of H-1 PAR- and PML-bodies by double-fluorescence labeling and confocal microscopy and found them to be spatially unrelated. Furthermore, H-1 PAR-bodies did not colocalize with other prominent nuclear structures such as nucleoli, coiled bodies, and speckled domains. Electron microscopy analysis revealed that NS1, as detected by indirect immunogold labeling, was localized in ring-shaped electron-dense nuclear structures corresponding in size and frequency to H-1 PAR-bodies. These structures were also clearly visible without immunogold labeling and could be detected only in infected cells. Our results suggest that H-1 virus does not target known nuclear bodies for DNA replication but rather induces the formation of a novel structure in the nucleus of infected cells.

* Corresponding author. Mailing address: Applied Tumor Virology Unit, Abteilung F0100 and INSERM U 375, Deutsches Krebsforschungszentrum, Postfach 101949, 69009 Heidelberg, Germany. Phone: 49 6221 424973. Fax: 49 6221 424962. E-mail: C.Cziepluch{at}dkfz-heidelberg.de.

Journal of Virology, May 2000, p. 4807-4815, Vol. 74, No. 10
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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