title: Early detection of a two-long-terminal-repeat junction molecule in the cytoplasm of recombinant murine leukemia virus-infected cells.
abstract: We showed that a U5-U3 junction was reproducibly detected by a PCR assay as early as 1 to 2 h postinfection with a DNase-treated murine leukemia virus (MLV)-containing supernatant in aphidicolin-arrested NIH 3T3 cells, as well as in nonarrested cells. Such detection is azidothymidine sensitive and corresponded to neosynthesized products of the reverse transcriptase. This observation was confirmed in two additional human cell lines, TE671 and ARPE-19. Using cell fractionation combined with careful controls, we found that a two-long-terminal-repeat (two-LTR) junction molecule was detectable in the cytoplasm as early as 2 h post virus entry. Altogether, our data indicated that the neosynthesized retroviral DNA led to the early formation of structures including true two-LTR junctions in the cytoplasm of MLV-infected cells. Thus, the classical assumption that two-LTR circles are a mitosis-dependent dead-end product accumulating in the nucleus must be reconsidered. MLV-derived products containing a two-LTR junction can no longer be used as an exclusive surrogate for the preintegration complex nuclear translocation event.
authors.affiliation: INSERM ERM 0-105, CHU Hôtel-Dieu, 30 blvd. Jean Monnet, 44035 Nantes Cedex 01, France.
authors.forename: Fatima
authors.initials: F
authors.lastname: Serhan
authors.forename: Magalie
authors.initials: M
authors.lastname: Penaud
authors.forename: Caroline
authors.initials: C
authors.lastname: Petit
authors.forename: Thierry
authors.initials: T
authors.lastname: Leste-Lasserre
authors.forename: Stéphane
authors.initials: S
authors.lastname: Trajcevski
authors.forename: David
authors.initials: D
authors.lastname: Klatzmann
authors.forename: Ghislaine
authors.initials: G
authors.lastname: Duisit
authors.forename: Pierre
authors.initials: P
authors.lastname: Sonigo
authors.forename: Philippe
authors.initials: P
authors.lastname: Moullier
mesh_terms: D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D003593:Cytoplasm / Q000235:genetics* / Q000821:virology; D004279:DNA, Viral / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005822:Genetic Vectors; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D051379:Mice; D008938:Mitosis; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells / Q000187:drug effects; D011995:Recombination, Genetic; D020079:Terminal Repeat Sequences / Q000235:genetics*; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D004791:Enzyme Inhibitors; D016590:Aphidicolin