Sequence-specific interaction between HIV-1 matrix protein and viral genomic RNA revealed by in vitro genetic selection.
The human immunodeficiency virus type-1 matrix protein (HIV-1 MA) is a multifunctional structural protein synthesized as part of the Pr55 gag polyprotein. We have used in vitro genetic selection to identify an RNA consensus sequence that specifically interacts with MA (Kd = 5 x 10(-7) M). This 13-nt MA binding consensus sequence bears a high degree of homology (77%) to a region (nt 1433-1446) within the POL open reading frame of the HIV-1 genome (consensus sequence from 38 HIV-1 strains). Chemical interference experiments identified the nucleotides within the MA binding consensus sequence involved in direct contact with MA. We further demonstrate that this RNA-protein interaction is mediated through a stretch of basic amino acids within MA. Mutations that disrupt the interaction between MA and its RNA binding site within the HIV-1 genome resulted in a measurable decrease in viral replication.
Purohit
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P
Program in Gene Function and Expression, Howard Hughes Medical Institute, University of Massachusetts Medical Center, Worcester 01605, USA.
Dupont
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Stevenson
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Green
M R
MR
D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D019020:Directed Molecular Evolution; D015683:Gene Products, gag / Q000378:metabolism*; D015684:Gene Products, pol / Q000235:genetics; D015488:HIV Antigens / Q000378:metabolism*; D015497:HIV-1 / Q000254:growth & development*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011498:Protein Precursors / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D013601:T-Lymphocytes / Q000821:virology; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D015684:Gene Products, pol; D015488:HIV Antigens; D011498:Protein Precursors; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C054858:p17 protein, Human Immunodeficiency Virus Type 1; C059767:p55 gag precursor protein, Human immunodeficiency virus 1