Efficient intracellular retrotransposition of an exogenous primate retrovirus genome.
The foamy virus (FV) subgroup of Retroviridae reverse transcribe their RNA (pre-)genome late in the replication cycle before leaving an infected cell. We studied whether a marker gene-transducing FV vector is able to shuttle to the nucleus and integrate into host cell genomic DNA. While a potential intracellular retrotransposition of vectors derived from other retroviruses was below the detection limit of our assay, we found that up to 5% of cells transfected with the FV vector were stably transduced, harboring 1 to approximately 10 vector integrants. Generation of the integrants depended on expression of functional capsid, reverse transcriptase and integrase proteins, and did not involve an extracellular step. PCR analysis of the U3 region of the 5' long terminal repeat and determination of proviral integration sites showed that a reverse transcription step had taken place to generate the integrants. Co-expression of a mutated envelope allowing particle egress and avoiding extracellular infection resulted in a significantly increased rescue of cells harboring integrants, suggesting that accumulation of proviruses via intracellular retrotransposition represents an integral part of the FV replication strategy.
Heinkelein
M
M
Institut für Virologie und Immunbiologie, Universität Würzburg and Institut für Virologie, Medizinische Fakultät 'Carl Gustav Carus', Technische Universität Dresden, Gerichtsstrasse 5, 01069 Dresden, Germany.
Pietschmann
T
T
Jármy
G
G
Dressler
M
M
Imrich
H
H
Thurow
J
J
Lindemann
D
D
Bock
M
M
Moebes
A
A
Roy
J
J
Herchenröder
O
O
Rethwilm
A
A
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D015686:Gene Products, env / Q000235:genetics; D005822:Genetic Vectors; D016679:Genome, Viral; D006367:HeLa Cells; D006801:Humans; D011323:Primates / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D018626:Retroelements; D016092:Spumavirus / Q000235:genetics*
D017931:DNA Primers; D015686:Gene Products, env; D012367:RNA, Viral; D018626:Retroelements