Human T-cell leukemia virus (HTLV) type II Rex protein binds specifically to RNA sequences of the HTLV long terminal repeat but poorly to the human immunodeficiency virus type 1 Rev-responsive element.
The human T-cell leukemia viruses (HTLVs) encode a trans-regulatory protein, Rex, which differentially regulates viral gene expression by controlling the cytoplasmic accumulation of viral mRNAs. Because of insufficient amounts of purified protein, biochemical characterization of Rex activity has not previously been performed. Here, utilizing the baculovirus expression system, we purified HTLV type II (HTLV-II) Rex from the cytoplasmic fraction of recombinant baculovirus-infected insect cells by heparin-agarose chromatography. We directly demonstrated that Rex specifically bound HTLV-II 5' long terminal repeat RNA in both gel mobility shift and immunobinding assays. Sequences sufficient for Rex binding were localized to the R-U5 region of the HTLV-II 5' long terminal repeat and correlate with the region required for Rex function. The human immunodeficiency virus type 1 (HIV-1), has an analogous regulatory protein, Rev, which directly binds to and mediates its action through the Rev-responsive element located within the HIV-1 env gene. We demonstrated that HTLV-II Rex rescued an HIV-1JR-CSF Rev-deficient mutant, although inefficiently. This result is consistent with a weak binding activity to the HIV-1 Rev-responsive element under conditions in which it efficiently bound the HTLV-II long terminal repeat RNA.
Yip
M T
MT
Department of Microbiology and Immunology, University of California, School of Medicine, Los Angeles 90024-1678.
Dynan
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Green
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Black
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Arrigo
S J
SJ
Torbati
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Heaphy
S
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Ruland
C
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Rosenblatt
J D
JD
Chen
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D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D015694:Gene Products, rev / Q000378:metabolism*; D016370:Gene Products, rex / Q000302:isolation & purification / Q000378:metabolism*; D015752:Genes, env; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D015367:Human T-lymphotropic virus 2 / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D016370:Gene Products, rex; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus