A plant virus replication system to assay the formation of RNA pseudotriloop motifs in RNA-protein interactions.
A pseudotriloop is formed by transloop base pairing between the first (5') and the fifth nucleotide in a hexanucleotide RNA loop ("hexaloop") to subtend a triloop of nucleotides 2-4. This structure has been found in hairpins involved in the regulation of iron metabolism in mammalian cells and in transcription of plant virus subgenomic RNA. Several hexaloop hairpins, including HIV-transactivation-responsive element and hepatitis B virus , potentially adopt a pseudotriloop conformation. Here we show that an RNA plant virus whose replication depends on a conventional triloop hairpin can be used to verify the existence of pseudotriloop structures in vivo. Our data suggest that the pseudotriloop may represent a common motif in RNA-protein recognition.
Haasnoot
P C Joost
PC
Institute of Biology, Gorlaeus Laboratories, Leiden University, 2333 CC Leiden, The Netherlands.
Bol
John F
JF
Olsthoorn
René C L
RC
D017794:Alfalfa mosaic virus / Q000235:genetics / Q000502:physiology; D001483:Base Sequence; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010942:Plant Viruses / Q000235:genetics* / Q000502:physiology; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics; D014779:Virus Replication
D012367:RNA, Viral; D014764:Viral Proteins