JVI Accepts, published online ahead of print on 2 July 2008

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J. Virol. doi:10.1128/JVI.00530-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Functional Analysis of the Complex TAR RNA Structure in Simian Immunodeficiency Virus

Mireille Centlivre, Bep Klaver, Ben Berkhout, and Atze T. Das^*

Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center of the University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands

^* To whom correspondence should be addressed. Email: a.t.das{at}amc.uva.nl.

Transcription of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) is activated through binding of the viral Tat protein to the trans-activating response (TAR) element at the 5' end of the nascent transcript. Whereas HIV-1 TAR folds a simple hairpin structure, the corresponding domain of HIV-2 and SIVmac exhibits a more complex structure composed of three stem-loops. This structural polymorphism may be attributed to additional functions of TAR in HIV-2/SIVmac replication. We recently constructed an SIVmac variant that does not require the Tat-TAR interaction for transcription. We used this variant to study additional roles of TAR in SIVmac replication and generated mutants with a truncated TAR structure. We demonstrate that partial or nearly complete removal of TAR does not impair viral transcription, RNA processing and translation. Moreover, these deletions do not significantly affect virus replication in the PM1 T cell line and macaque peripheral blood mononuclear cells (PBMC). These results demonstrate that the complex TAR structure in SIVmac has no other essential function in in vitro virus replication besides its role in Tat-mediated activation of transcription.