Stimulation of Tat-independent transcriptional processivity from the HIV-1 LTR promoter by matrix attachment regions.
The chromatin environment and the sites of integration in the host genome are critical determinants of human immunodeficiency virus (HIV) transcription and replication. Depending on the chromosomal location of provirus integration within the genome, HIV-1 long terminal repeat (LTR)-mediated transcription may vary from 0- to 70-fold. Cis-elements such as topoisomerase II cleavage sites, Alu repeats and matrix attachment regions (MARs) are thought to be targets for retroviral integration. Here we show that a novel MAR sequence from the T-cell receptor beta locus (MARbeta) and the IgH MAR mediate transcriptional augmentation when placed upstream of the HIV-1 LTR promoter. The effect of transcriptional augmentation is seen in both transient and stable transfection, indicating its effect even upon integration in the genome. MAR-mediated transcriptional elevation is independent of Tat, and occurs synergistically in the presence of Tat. Further, we show that MAR-mediated transcriptional elevation is specific to the HIV-1 LTR and the Moloney murine leukemia virus LTR promoter. In a transient transfection assay using over-expressed IkappaB, the inhibitor of NF-kappaB, we show that MAR-induced processive transcription is NF-kappaB dependent, signifying the role of local enhancers within the LTR promoter. Furthermore, by RNase protection experiments using proximal and distal probes, we show that MAR-mediated transcriptional upregulation is more prominent at the distal rather than the proximal end, thus indicating the potential role of MARs in promoting elongation.
Rampalli
Shravanti
S
National Center for Cell Science, Pune University Campus, Ganeshkhind, Pune 411007, India.
Kulkarni
Asavari
A
Kumar
Pavan
P
Mogare
Devraj
D
Galande
Sanjeev
S
Mitra
Debashis
D
Chattopadhyay
Samit
S
D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D015967:Gene Expression Regulation, Viral; D015696:Gene Products, tat / Q000502:physiology; D019674:Genes, T-Cell Receptor beta; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D016328:NF-kappa B / Q000502:physiology; D015530:Nuclear Matrix / Q000378:metabolism; D010441:Peptide Chain Elongation, Translational; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D015533:Transcriptional Activation; D016662:Virus Integration; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D007143:Immunoglobulin Heavy Chains; D016328:NF-kappa B; D054322:tat Gene Products, Human Immunodeficiency Virus