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HIV-1 provirus, either as a chromosomal integrant or as an episomal plasmid in HeLa cells, forms a transcription-dependent gene loop structure between the 5'LTR promoter and 3'LTR poly(A) signal. Flavopiridol-mediated inhibition of RNA polymerase II elongation blocks 5' to 3'LTR juxtaposition, indicating that this structure is maintained during transcription. Analysis of mutant or hybrid HIV-1 plasmids demonstrates that replacement of the 5'LTR promoter with CMV or the 3'LTR poly(A) signal with a synthetic element (SPA) permits gene loop formation, suggesting that these interactions are not retroviral specific. In addition, activation of the 5'LTR poly(A) signal or inactivation of the 3'LTR poly(A) signal abolishes gene loop formation. Overall, we demonstrate that both ongoing transcription and pre-mRNA processing are essential for gene loop formation, and predict that these structures represent a defining feature of active gene transcription.

Original publication

DOI

10.1016/j.molcel.2007.11.030

Type

Journal article

Journal

Mol Cell

Publication Date

18/01/2008

Volume

29

Pages

56 - 68

Keywords

Chromatin, Chromatin Immunoprecipitation, Cytomegalovirus, Defective Viruses, Flavonoids, Gene Expression Regulation, Viral, Genes, tat, HIV Long Terminal Repeat, HIV-1, HeLa Cells, Humans, Nucleic Acid Conformation, Phosphoserine, Piperidines, Promoter Regions, Genetic, Proviruses, RNA Polymerase II, RNA Precursors, RNA Processing, Post-Transcriptional, RNA, Messenger, Transcription, Genetic, Transfection, U937 Cells