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J. Biol. Chem., Vol. 276, Issue 52, 48803-48813, December 28, 2001
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From the 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 The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF281068.
Organization of the Human tarbp2 Gene Reveals Two
Promoters That Are Repressed in an Astrocytic Cell Line*
§,,
,,§§, and¶§§¶¶
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 Department of
Microbiology and Immunology, University of Melbourne,
Parkville 3010, Victoria, Australia
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.
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.
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