DNA, Viral; Luminescent Proteins; Green Fluorescent Proteins; Base Sequence; CpG Islands; DNA Methylation; DNA, Viral/genetics; DNA, Viral/metabolism; Genetic Vectors; HIV Long Terminal Repeat; HIV-1/genetics; HIV-1/metabolism; Humans; Immunologic Memory; In Vitro Techniques; Jurkat Cells; Luminescent Proteins/genetics; Promoter Regions, Genetic; Proviruses/genetics; Proviruses/metabolism; Suppression, Genetic; T-Lymphocytes/immunology; Transduction, Genetic; Virus Activation/genetics; Virus Integration/genetics; Microbiology; Immunology; Insect Science; Virology
Abstract :
[en] Persistence of human immunodeficiency virus type 1 (HIV-1) constitutes a major obstacle in the control of HIV-1 infection. Here we investigated whether the CpG methylation of the HIV-1 promoter can directly influence the expression of the HIV-1 genome and thereby contribute to the persistence and latency of HIV-1. The levels of CpG methylation in the promoter of HIV-1 were studied after bisulfite-induced modification of DNA in five Jurkat clonal cell lines transduced by an HIV-1 long terminal repeat (LTR)-driven retroviral vector and expressing enhanced green fluorescent protein (GFP) and in primary resting memory T cells challenged with HIV-1 or with an HIV-1-derived retroviral vector. Basal HIV-1 promoter activities were low or undetectable in three tested HIV-1 LTR-GFP clones, one of which encoded the Tat protein, and they reached medium or high levels in two other clones. The CpG dinucleotide that occurred in a latently infected clonal cell line 240 nucleotides upstream of the transcription start remained methylated after reactivation of HIV-1 transcription with 10 nM phorbol-12-myristate-13-acetate. In two clones showing a medium promoter activity and in resting memory T cells, the HIV-1 LTR was generally not methylated. Our results show that the methylation profiles of the HIV-1 LTR, including those present in latently infected cells, are low and do not correlate with the transcriptional activity. We suggest that, in a noncloned cellular population in which the HIV-1 proviruses are randomly integrated in the human genome, HIV-1 latency is imperfectly controlled by CpG methylation and is inherently accompanied by residual replication.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Pion, Marjorie; INSERM U372, Unité de Pathogénie des Infections à Lentivirus, Parc Scientifique et Technologique de Luminy, 13276 Marseille, France
Jordan, Albert; Gladstone Inst. of Virol./Immunology, San Francisco, CA 941412, United States
Biancotto, Angelique; INSERM U372, U. Pathog. des Infect. a Lentivirus, Parc Sci. et Technologique de Luminy, 13276 Marseille, France
Dequiedt, Franck ; Université de Liège - ULiège > Département des sciences de la vie > Génétique et biologie moléculaires animales ; Gladstone Inst. of Virol./Immunology, San Francisco, CA 941412, United States
Gondois-Rey, Françoise; INSERM U372, U. Pathog. des Infect. a Lentivirus, Parc Sci. et Technologique de Luminy, 13276 Marseille, France
Rondeau, Sophie; INSERM U372, U. Pathog. des Infect. a Lentivirus, Parc Sci. et Technologique de Luminy, 13276 Marseille, France
Vigne, Robert; INSERM U372, U. Pathog. des Infect. a Lentivirus, Parc Sci. et Technologique de Luminy, 13276 Marseille, France
Hejnar, Jiri; Institute of Molecular Genetics, Acad. of Sci. of the Czech Republic, 166 37 Prague, Czech Republic
Verdin, Eric; Gladstone Inst. of Virol./Immunology, San Francisco, CA 941412, United States
Hirsch, Ivan; INSERM U372, U. Pathog. des Infect. a Lentivirus, Parc Sci. et Technologique de Luminy, 13276 Marseille, France ; INSERM U372, Lab. de Pathog. Infect. a Lentivirus, 13276 Marseille Cedex 9, France
Language :
English
Title :
Transcriptional suppression of in vitro-integrated human immunodeficiency virus type 1 does not correlate with proviral DNA methylation.
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