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Originally published In Press as doi:10.1074/jbc.M104645200 on October 18, 2001

J. Biol. Chem., Vol. 276, Issue 52, 48803-48813, December 28, 2001
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Organization of the Human tarbp2 Gene Reveals Two Promoters That Are Repressed in an Astrocytic Cell Line*

Sylvie BannwarthDagger §, Lily TalakoubDagger , Franck Letourneur, Mariela Duarte||, Damian F. Purcell**Dagger Dagger , John HiscottDagger §§, and Anne GatignolDagger §§¶¶

From the Dagger  Molecular Oncology Group, McGill AIDS Centre, Lady Davis Institute for Medical Research and the §§ Departments of Medicine and Microbiology & Immunology, McGill University, Montreal, Quebec H3T 1E2, Canada,  INSERM, Institut Cochin de Génétique Moléculaire, 75014 Paris, France, the ** Macfarlane Burnet Centre for Medical Research, Fairfield 3078, Victoria, Australia, and the Dagger Dagger  Department of Microbiology and Immunology, University of Melbourne, Parkville 3010, Victoria, Australia

TRBP1 and TRBP2 are isoforms of a double-stranded RNA-binding protein that differ in their N-terminal end and were each identified by binding to human immunodeficiency virus type 1 (HIV-1) trans-activation-responsive RNA. TRBP1 and TRBP2 also bind and modulate the function of the double-stranded RNA-activated protein kinase, protein kinase R. Both proteins increase long terminal repeat expression in human and murine cells, and their gene has been mapped to human chromosome 12. We have isolated and characterized the complete tarbp2 gene (5493 bp) coding for the two TRBP proteins. Two adjacent promoters initiate transcription of alternative first exons for TRBP1 and TRBP2 mRNAs that are spliced onto common downstream exons. TRBP2 transcription and translation start sites are localized within the first intron of TRBP1. TRBP promoters are TATA-less but have CCAAT boxes, a CpG island, and several potential binding sites for transcriptional factors. Promoter deletion analysis identified two regions from position -1397 to -330 for TRBP1 and from position -330 to +38 for TRBP2 that are important for promoter function. TRBP2 promoter activity was expressed at a higher level compared with TRBP1 promoter. In addition, a specific down-regulation of TRBP1 and TRBP2 promoter activity was identified in human astrocytic cell line U251MG compared with HeLa cells. This minimal TRBP promoter activity may account for minimal HIV-1 replication in astrocytes.

* This work was supported by grants from the Agence Nationale de Recherches sur le SIDA, Medical Research Council Grant 38112, Canadian Foundation for AIDS Research Grant 013513 (to A. G.), Grant 111700 from the National Health and Medical Research Council of Australia (to D. F. P.), and by a Medical Research Council program grant (to J. H.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF281068.

§ Supported by a fellowship from the Fondation pour la Recherche Médicale, France.

|| Supported by a fellowship from Ensemble contre le SIDA, France. Present address: CEA-LGRK, 2 rue Gaston Crémieux, BP 22, 91057 Evry Cédex, France.

¶¶ Recipient of an INSERM, France/MRC, Canada award. Research Scientist from the Fond de la Recherche en Santé du Québec. To whom correspondence should be addressed: Molecular Oncology Group, McGill AIDS Center, Lady Davis Institute for Medical Research, 3755 Côte Ste. Catherine, Montréal, QC H3T 1E2, Canada. Tel.: 514-340-8260, Ext. 5284; Fax: 514-340-7576; E-mail: anne.gatignol@mcgill.ca.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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