JVI Accepts, published online ahead of print on 3 June 2009

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J. Virol. doi:10.1128/JVI.00554-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Functional characterization of Negri bodies (NBs) in rabies virus infected cells: evidence that NBs are sites of viral transcription and replication

Xavier Lahaye, Aurore Vidy, Carole Pomier, Linda Obiang, Francis Harper, Yves Gaudin, and Danielle Blondel^*

CNRS, UMR2472, INRA, UMR1157, IFR 115, Virologie Moléculaire et Structurale, 91198, Gif sur Yvette, France; CNRS, FRE 2937, IFR 89, Laboratoire de Replication de l'ADN et Ultrastructure du noyau, 94801 Villejuif, France

^* To whom correspondence should be addressed. Email: blondel{at}vms.cnrs-gif.fr.

Rabies virus infection induces the formation of cytoplasmic inclusion bodies that resemble Negri bodies found in the cytoplasm of some infected nerve cells. We have studied the morphogenesis and the role of these Negri bodies-like structures (NBLs) during viral infection. The results indicate that these spherical structures (1 or 2 per cell in the initial stage of infection), composed of the viral N and P proteins, grow during the virus cycle before appearing as smaller structures at late stages of infection. We have shown that the microtubule network is not necessary for the formation of these inclusion bodies but is involved in their dynamics. In contrast, the actin network does not play any detectable role in these processes. These inclusion bodies contain Hsp70 and ubiquitinylated proteins but they are not misfolded proteins aggregates. NBLs in fact appear to be functional structures involved in the viral life cycle. Specifically, using in situ fluorescent hybridization techniques (FISH), we have shown that all viral RNAs (genome, antigenome and every messenger RNA) are located inside the inclusion bodies. Significantly, short-term RNA labeling in presence of BrUTP strongly suggests that the NBLs are the sites where viral transcription and replication take place.

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