title: Construction of a reporter vector for analysis of bidirectional transcriptional activity of retrovirus LTR.
abstract: To study the transcriptional activity of the HIV-1 LTR, we constructed a vector containing Renilla and Firefly luciferase genes under the control of the LTR (wild-type or mutated version) and oriented in a manner that allowed them to be transcribed in opposite directions. We found that the HIV-1 LTR acted as a bidirectional promoter, which activity was controlled by NF-κB- and Sp1-binding sites in both orientations. We next analyzed with this reporter vector the bidirectional promoter activity of the HTLV-1 LTR and showed that this LTR also possessed a bidirectional transcriptional activity. Interestingly, Sp1-binding elements were also involved in the control of HTLV-1 bidirectional transcription. Moreover, both retroviral trans-activators, Tat and Tax, could preferentially activate sense transcription with no or limited effect on the extent of antisense transcription. We also cloned into this plasmid the MLV LTR and found that the LTR of a simple retrovirus also possessed bidirectional transcriptional activity. This reporter vector represents a powerful tool to analyze the bidirectional transcriptional activity of retrovirus LTRs.
authors: Université Montpellier 1, Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé (CPBS), France; CNRS, UM5236, CPBS, F-34293 Montpellier, France.
authors: Charlotte
authors: C
authors: Arpin-André
authors: Université Montpellier 1, Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé (CPBS), France; CNRS, UM5236, CPBS, F-34293 Montpellier, France; East Carolina University, Department of Microbiology and Immunology, Greenville, NC 27834, USA.
authors: Sylvain
authors: S
authors: Laverdure
authors: Université du Québec à Montréal, Département des sciences biologiques and Centre de recherche BioMed, Montréal, Québec H2X 3X8, Canada.
authors: Benoit
authors: B
authors: Barbeau
authors: Université Montpellier 1, Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé (CPBS), France; CNRS, UM5236, CPBS, F-34293 Montpellier, France. Electronic address: antoine.gross@cpbs.cnrs.fr.
authors: Antoine
authors: A
authors: Gross
authors: Université Montpellier 1, Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé (CPBS), France; CNRS, UM5236, CPBS, F-34293 Montpellier, France. Electronic address: jean-michel.mesnard@cpbs.cnrs.fr.
authors: Jean-Michel
authors: JM
authors: Mesnard
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D015696:Gene Products, tat / Q000235:genetics; D016356:Gene Products, tax / Q000235:genetics; D017930:Genes, Reporter; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics*; D015368:Human T-lymphotropic virus 1 / Q000235:genetics*; D006801:Humans; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic; D020079:Terminal Repeat Sequences / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection
chemical_list: D015696:Gene Products, tat; D016356:Gene Products, tax; D016328:NF-kappa B
keywords: Antisense transcription; Bidirectional promoter; HIV-1; HTLV-1; Tat; Tax