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Journal of Virology, August 2009, p. 7948-7958, Vol. 83, No. 16
0022-538X/09/$08.00+0     doi:10.1128/JVI.00554-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Functional Characterization of Negri Bodies (NBs) in Rabies Virus-Infected Cells: Evidence that NBs Are Sites of Viral Transcription and Replication{triangledown}

Xavier Lahaye,1 Aurore Vidy,1,{dagger} Carole Pomier,1 Linda Obiang,1 Francis Harper,2 Yves Gaudin,1 and Danielle Blondel1*

CNRS, UMR2472, INRA, UMR1157, IFR 115, Virologie Moléculaire et Structurale, 91198, Gif sur Yvette, France,1 CCNRS, FRE 2937, IFR 89, Laboratoire de Replication de l'ADN et Ultrastructure du Noyau, 94801 Villejuif, France2

Received 18 March 2009/ Accepted 26 May 2009

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 body-like structures (NBLs) during viral infection. The results indicate that these spherical structures (one or two 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 protein aggregates. NBLs, in fact, appear to be functional structures involved in the viral life cycle. Specifically, using in situ fluorescent hybridization techniques, we show that all viral RNAs (genome, antigenome, and every mRNA) are located inside the inclusion bodies. Significantly, short-term RNA labeling in the presence of BrUTP strongly suggests that the NBLs are the sites where viral transcription and replication take place.

* Corresponding author. Mailing address: CNRS, UMR2472, INRA, UMR1157, IFR 115, Virologie Moléculaire et Structurale, 91198, Gif sur Yvette, France. Phone: 33 1 69 82 38 37. Fax: 33 1 69 82 43 08. E-mail: blondel{at}vms.cnrs-gif.fr

{triangledown} Published ahead of print on 3 June 2009.

{dagger} Present address: Max von Pettenkofer Institute für Virologie, Ludwig Maximilians University, Pettenkoferstrasse 9a, 80336 Munich, Germany.

Journal of Virology, August 2009, p. 7948-7958, Vol. 83, No. 16
0022-538X/09/$08.00+0     doi:10.1128/JVI.00554-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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