Effects of insertion of multiple AP-1 binding sites into the U3 region of the long terminal repeat of feline immunodeficiency virus.
An oligonucleotide containing multiple AP-1 binding sites was introduced into the regulatory sequence in the long terminal repeat (LTR) of feline immunodeficiency virus (FIV). Chloramphenicol acetyltransferase assay revealed that basal promoter activity of the mutated LTR was higher than that of the wild-type LTR in Crandell feline kidney (CRFK) cells. The mutated LTR was introduced into an infectious molecular clone of FIV and the clone was transfected into CRFK cells. The virus production of the mutant in the cells was as high as that of the wild-type when determined by the reverse transcriptase activity assay. The growth of the mutant virus obtained from the transfected CRFK cells was examined in feline T lymphoblastoid cell lines (MYA-1 and FeL-039 cells) and primary feline peripheral blood mononuclear cells (fPBMCs). The growth was delayed when compared with that of the wild-type virus in all the cells used. Upon examination by polymerase chain reaction, the length of the LTR of the mutant virus was shortened in both MYA-1 cells and fPBMCs. Sequence analysis revealed that the insertion was completely deleted 39 days after infection in the MYA-1 cells.
Miyazawa
T
T
Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.
Tomonaga
K
K
Kawaguchi
Y
Y
Kohmoto
M
M
Inoshima
Y
Y
Maeda
K
K
Mikami
T
T
Maedadel K [corrected to Maeda
K ]
K]
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002415:Cats; D002460:Cell Line; D017931:DNA Primers; D005822:Genetic Vectors; D016089:Immunodeficiency Virus, Feline / Q000235:genetics* / Q000502:physiology; D007668:Kidney; D007700:Kinetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D014779:Virus Replication
D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D018808:Transcription Factor AP-1; D012194:RNA-Directed DNA Polymerase