Transcription-dependent gene looping of the HIV-1 provirus is dictated by recognition of pre-mRNA processing signals.
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.
Perkins
Kelly J
KJ
Sir William Dunn School of Pathology, South Parks Road, University of Oxford, Oxford OX1 3RE, UK.
Lusic
Marina
M
Mitar
Ivonne
I
Giacca
Mauro
M
Proudfoot
Nick J
NJ
D002843:Chromatin / Q000235:genetics / Q000648:ultrastructure*; D047369:Chromatin Immunoprecipitation; D003587:Cytomegalovirus / Q000235:genetics; D003673:Defective Viruses / Q000235:genetics / Q000502:physiology; D005419:Flavonoids / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral; D015756:Genes, tat; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D009690:Nucleic Acid Conformation; D010768:Phosphoserine / Q000032:analysis; D010880:Piperidines / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011533:Proviruses / Q000648:ultrastructure*; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D012322:RNA Precursors / Q000737:chemistry / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection; D020298:U937 Cells / Q000821:virology
D002843:Chromatin; D005419:Flavonoids; D010880:Piperidines; D012322:RNA Precursors; D012333:RNA, Messenger; D010768:Phosphoserine; C077990:alvocidib; D012319:RNA Polymerase II