Functional Differences between the Long Terminal Repeat Transcriptional Promoters of Human Immunodeficiency Virus Type 1 Subtypes A through G

doi:10.1128/JVI.74.8.3740-3751.2000

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Journal of Virology, April 2000, p. 3740-3751, Vol. 74, No. 8
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Functional Differences between the Long Terminal Repeat Transcriptional Promoters of Human Immunodeficiency Virus Type 1 Subtypes A through G

Rienk E. Jeeninga, Maarten Hoogenkamp, Mercedes Armand-Ugon, Michel de Baar, Koen Verhoef, and Ben Berkhout^*

Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

Received 24 September 1999/Accepted 25 January 2000

The current human immunodeficiency virus type 1 (HIV-1) shows an increasing number of distinct viral subtypes, as well asviruses that are recombinants of at least two subtypes. Althoughno biological differences have been described so far for virusesthat belong to different subtypes, there is considerable sequencevariation between the different HIV-1 subtypes. The HIV-1 longterminal repeat (LTR) encodes the transcriptional promoter, andthe LTR of subtypes A through G was cloned and analyzed to testif there are subtype-specific differences in gene expression.Sequence analysis demonstrated a unique LTR enhancer-promoterconfiguration for each subtype. Transcription assays with luciferasereporter constructs showed that all subtype LTRs are functionalpromoters with a low basal transcriptional activity and a highactivity in the presence of the viral Tat transcriptional activatorprotein. All subtype LTRs responded equally well to the Tat transactivator protein of subtype B. This result suggests that thereare no major differences in the mechanism of Tat-mediated transactivation among the subtypes. Nevertheless, subtype-specificdifferences in the activity of the basal LTR promoter were measuredin different cell types. Furthermore, we measured a differentialresponse to tumor necrosis factor alpha treatment, and the inductionlevel correlated with the number of NF- $kappa$ B sites in the respectiveLTRs, which varies from one (subtype E) to three (subtype C).In general, subtype E was found to encode the most potent LTR,and we therefore inserted the core promoter elements of subtypeE in the infectious molecular clone of the LAI isolate (subtypeB). This recombinant LAI-E virus exhibited a profound replicationadvantage compared with the original LAI virus in the SupT1 T-cellline, indicating that subtle differences in LTR promoter activitycan have a significant impact on viral replication kinetics. Theseresults suggest that there may be considerable biological differencesamong the HIV-1subtypes.

^* Corresponding author. Mailing address: Department of Human Retrovirology, Academic Medical Center, University of Amsterdam,Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: (31-20)566 4822. Fax: (31-20) 691 6531. E-mail: b.berkhout{at}amc.uva.nl.

Journal of Virology, April 2000, p. 3740-3751, Vol. 74, No. 8
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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