Specific binding of polypyrimidine tract binding protein and hnRNP A1 to HIV-1 CRS elements.
The human immunodeficiency virus (HIV) Rev and human T-cell leukemia virus (HTLV) Rex proteins regulate viral RNA processing. Both proteins act to overcome the block to viral structural gene expression, at least in part, by reversing the inhibitory effect of intronic RNA sequences, termed cis-acting repressive (CRS) sequences. Using HTLV type II (HTLV-II) as a model, we recently showed that the function of a 5' long terminal repeat (LTR) CRS correlates with in vitro binding by both polypyrimidine tract binding (PTB) protein (also known as hnRNP I) and hnRNP A1 to CRS RNA (1,2). Using radioimmunoprecipitation of proteins ultraviolet (UV) crosslinked to each HIV CRS RNA with monoclonal anti-hnRNP antibodies, we now demonstrate that hnRNP I and hnRNP A1 bind to two different HIV-1 CRS RNAs. In addition, we show that hnRNP I and hnRNP A1 binding to HIV-1 CRS RNAs can be specifically competed by HTLV-II CRS RNAs using electrophoretic mobility shift assay (EMSA)/UV crosslinking assays. Binding by both hnRNP I and hnRNP A1 to HIV-1 and HTLV-II CRS RNAs suggests a role for these proteins in CRS function that may be influenced by the Rev and Rex proteins, respectively.
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Luo
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Bakker
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Fraser
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Rosenblatt
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D004591:Electrophoresis, Polyacrylamide Gel; D015685:Fusion Proteins, gag-pol / Q000378:metabolism; D015694:Gene Products, rev / Q000378:metabolism*; D016370:Gene Products, rex / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D015367:Human T-lymphotropic virus 2 / Q000235:genetics; D006801:Humans; D038941:Polypyrimidine Tract-Binding Protein; D015347:RNA Probes / Q000378:metabolism; D012332:RNA, Heterogeneous Nuclear / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D054321:rev Gene Products, Human Immunodeficiency Virus
D015685:Fusion Proteins, gag-pol; D015694:Gene Products, rev; D016370:Gene Products, rex; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D015347:RNA Probes; D012332:RNA, Heterogeneous Nuclear; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D038941:Polypyrimidine Tract-Binding Protein