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J Virol, March 1998, p. 2305-2309, Vol. 72, No. 3
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

An Endonuclease Switching Mechanism in the Virion RNA and cRNA Promoters of Thogoto Orthomyxovirus

Michael B. Leahy,1 Johannes T. Dessens,1 David C. Pritlove,2 and Patricia A. Nuttall1,*

NERC Institute of Virology and Environmental Microbiology, Oxford OX1 3SR,1 and Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE,2 United Kingdom

Received 8 August 1997/Accepted 17 November 1997

An in vitro assay was developed to investigate endonuclease activity of Thogoto virus, a tick-borne orthomyxovirus. Endonuclease activity relied on an interaction between the 3' and 5' termini of virion RNA (vRNA) and not those of cRNA. Evidence was obtained that cap structures are cleaved directly from cap donors and that cleavage does not occur after pyrimidines. A 5' hook structure, present in the vRNA promoter but not the cRNA promoter, was introduced into cRNA promoter mutants. These mutants stimulated endonuclease activity, although at levels slightly lower than that of vRNA. The ability of the cRNA promoter to stimulate endonuclease activity when mutated to contain a 5' hook structure indicates that this structure constitutes a switching mechanism for endonuclease activity between the vRNA and cRNA promoters.

* Corresponding author. Mailing address: NERC Institute of Virology and Environmental Microbiology, Mansfield Road, Oxford OX1 3SR, United Kingdom. Phone: 44-1865-281631. Fax: 44-1865-281696. E-mail address: pan{at}mail.nerc-oxford.ac.uk.




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