Early detection of a two-long-terminal-repeat junction molecule in the cytoplasm of recombinant murine leukemia virus-infected cells.
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.
Serhan
Fatima
F
INSERM ERM 0-105, CHU Hôtel-Dieu, 30 blvd. Jean Monnet, 44035 Nantes Cedex 01, France.
Penaud
Magalie
M
Petit
Caroline
C
Leste-Lasserre
Thierry
T
Trajcevski
Stéphane
S
Klatzmann
David
D
Duisit
Ghislaine
G
Sonigo
Pierre
P
Moullier
Philippe
P
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
D004279:DNA, Viral; D004791:Enzyme Inhibitors; D016590:Aphidicolin