Identification of a high-affinity RNA-binding site for the human immunodeficiency virus type 1 Rev protein.
Expression of the structural proteins of human immunodeficiency virus type 1 requires the direct interaction of multiple copies of the viral Rev protein with its highly structured RNA target sequence, the Rev response element (RRE). Nucleotides critical for Rev monomer binding have been mapped by chemical interference to a single site flanking the base of an RNA helix (stem IIB) located within the 234-nucleotide RRE. Binding of additional Rev molecules to an RRE probe did not require any RNA primary sequence information detectable by modification interference beyond that required for binding of a single Rev protein molecule. A synthetic 29-nucleotide RNA molecule designed to incorporate nucleotides identified as critical for Rev binding retained the ability to bind Rev specifically and, therefore, represents a minimal Rev-binding site. We propose that Rev binding to the RRE initiates with the direct interaction of a Rev monomer with a high-affinity binding site located at the base of the IIB stem of the RRE. The subsequent formation of Rev multimers on the RRE appears, in contrast, primarily driven by specific protein-protein interactions.
Tiley
L S
LS
Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710.
Malim
M H
MH
Tewary
H K
HK
Stockley
P G
PG
Cullen
B R
BR
D001483:Base Sequence; D001665:Binding Sites; D004252:DNA Mutational Analysis; D015694:Gene Products, rev / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D012045:Regulatory Sequences, Nucleic Acid; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus