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Journal of Virology, June 2009, p. 5846-5853, Vol. 83, No. 11
0022-538X/09/$08.00+0     doi:10.1128/JVI.02602-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identification of an Intercistronic Internal Ribosome Entry Site in a Marek's Disease Virus Immediate-Early Gene{triangledown} ,{dagger}

Abdessamad Tahiri-Alaoui,1* Lorraine P. Smith,1 Suzan Baigent,1 Lydia Kgosana,1 Lawrence J. Petherbridge,1 Luke S. Lambeth,1 William James,2 and Venugopal Nair1*

Institute for Animal Health, Division of Microbiology, Compton, Berkshire RG20 7NN, United Kingdom,1 Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom2

Received 17 December 2008/ Accepted 10 March 2009

In this study, we have identified an internal ribosome entry site (IRES) from the highly infectious herpesvirus Marek's disease virus (MDV). The IRES was mapped to the intercistronic region (ICR) of a bicistronic mRNA that we cloned from the MDV-transformed CD4+ T-cell line MSB-1. The transcript is a member of a family of mRNAs expressed as immediate-early genes with two open reading frames (ORF). The first ORF encodes a 14-kDa polypeptide with two N-terminal splice variants, whereas the second ORF is contained entirely within a single exon and encodes a 12-kDa protein also known as RLORF9. We have shown that the ICR that separates the two ORFs functions as an IRES that controls the translation of RLORF9 when cap-dependent translation is inhibited. Deletion analysis revealed that there are two potential IRES elements within the ICR. Reverse genetic experiments with the oncogenic strain of MDV type 1 indicated that deletion of IRES-controlled RLORF9 does not significantly affect viral replication or MDV-induced mortality.

* Corresponding author. Mailing address: Institute for Animal Health, Division of Microbiology, Compton, Berkshire RG20 7NN, United Kingdom. Phone: 44 1635 577356. Fax: 44 1635 577263. E-mail for Abdessamad Tahiri-Alaoui: abdou.tahiri-alaoui{at}bbsrc.ac.uk. E-mail for Venugopal Nair: venu.gopal{at}bbsrc.ac.uk

{triangledown} Published ahead of print on 18 March 2009.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, June 2009, p. 5846-5853, Vol. 83, No. 11
0022-538X/09/$08.00+0     doi:10.1128/JVI.02602-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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