Affinity purification of a correctly folded fragment of synthetic HIV-1 mRNA using a HIV-1 Rev peptide-ligand.
Formation of a macromolecular complex between the RNA binding protein HIV-1 Rev and HIV-1 mRNA is an essential prerequisite for nuclear export and subsequent expression of HIV-1 mRNA. The arginine rich peptide TRQARRNRRRRWRARQR, corresponding to residues 34-50 of HIV-1 Rev, contains the mRNA binding motif. We prepared a thioether linked Rev34-50-cellulose conjugate to affinity purify a fragment of synthetic mRNA corresponding to the high affinity binding site for Rev. The correctly folded fraction of mRNA (27.5%) was isolated from a crude synthetic mixture.
Englebretsen
D R
DR
Centre for Drug Design and Development, University of Queensland, St. Lucia, Australia.
Scanlon
M J
MJ
West
M L
ML
D000595:Amino Acid Sequence; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods; D015694:Gene Products, rev / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics*; D008024:Ligands; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D011489:Protein Denaturation; D012333:RNA, Messenger / Q000737:chemistry / Q000302:isolation & purification*; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification*; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D008024:Ligands; D012333:RNA, Messenger; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus