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Journal of Virology, September 1999, p. 7153-7164, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Herpes Simplex Virus vhs Protein Induces Endoribonucleolytic Cleavage of Target RNAs in Cell Extracts

Mabrouk M. Elgadi,1 Christopher E. Hayes,1 and James R. Smiley2,3,*

Departments of Biology1 and Pathology,2 McMaster University, Hamilton, Ontario, Canada L8N 3Z5, and Department of Medical Microbiology & Immunology, University of Alberta, Edmonton, Alberta, Canada T6G 2H73

Received 19 March 1999/Accepted 24 May 1999

The herpes simplex virus virion host shutoff (vhs) protein (UL41 gene product) is a component of the HSV virion tegument that triggers shutoff of host protein synthesis and accelerated mRNA degradation during the early stages of HSV infection. Previous studies have demonstrated that extracts from HSV-infected cells and partially purified HSV virions display vhs-dependent RNase activity and that vhs is sufficient to trigger accelerated RNA degradation when expressed as the only HSV protein in an in vitro translation system derived from rabbit reticulocytes. We have used the rabbit reticulocyte translation system to characterize the mode of vhs-induced RNA decay in more detail. We report here that vhs-dependent RNA decay proceeds through endoribonucleolytic cleavage, is not affected by the presence of a 5' cap or a 3' poly(A) tail in the RNA substrate, requires Mg2+, and occurs in the absence of ribosomes. Intriguingly, sites of preferential initial cleavage were clustered over the 5' quadrant of one RNA substrate that was characterized in detail. The vhs homologue of pseudorabies virus also induced accelerated RNA decay in this in vitro system.

* Corresponding author. Mailing address: Department of Medical Microbiology & Immunology, 1-41, Medical Sciences Bldg., University of Alberta, Edmonton, Alberta, Canada T6G 2H7. Phone: (780) 492-2308. Fax: (780) 492-7521. E-mail: jim.smiley{at}ualberta.ca.

Journal of Virology, September 1999, p. 7153-7164, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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