Identity elements in bovine tRNA(Trp) required for the specific stimulation of gelonin, a plant ribosome-inactivating protein
| Autor: | Domenica Carnicelli, Simonetta Sperti, Simona Rizzi, Paola Accorsi, Alessandra Pallanca, Maurizio Brigotti, Lucio Montanaro |
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| Jazyk: | angličtina |
| Rok vydání: | 1999 |
| Předmět: |
Molecular Sequence Data
D arm Biology Ribosome medicine T7 RNA polymerase Animals Nucleotide Gelonin Molecular Biology Plant Proteins chemistry.chemical_classification Binding Sites Base Sequence RNA RNA Fungal RNA Transfer Trp Biochemistry chemistry Transfer RNA Ribosome Inactivating Proteins Type 1 Nucleic Acid Conformation Cattle T arm medicine.drug Research Article |
| Popis: | Ribosome-inactivating proteins (RIPs) are RNA-N-glycosidases widely present in plants that depurinate RNA in ribosomes at a specific universally conserved position, A4324, in the rat 28S rRNA. A small group of RIPs (cofactor-dependent RIPs) require ATP and tRNA to reach maximal activity on isolated ribosomes. Among cofactor-dependent RIPs, gelonin is specifically and uniquely stimulated by tRNA(Trp). The active species are avian (chicken) and mammalian (beef, rat, and rabbit) tRNA(Trp), whereas yeast tRNA(Trp) is completely devoid of stimulating activity. In the present article, bovine and yeast tRNA(Trp) with unmodified bases were prepared by assembly of the corresponding genes from synthetic oligonucleotides followed by PCR and T7 RNA polymerase transcription of the amplified products. The two synthetic tRNAs were fully active (bovine) or inactive (yeast) as the wild-type tRNAs. Construction of chimeric tRNA(Trp) transcripts identified the following bovine nucleotides as recognition elements for gelonin-stimulating activity: G26 and bp G12-C23 in the D arm and G57, A59, and bp G51-C63 and U52-A62 in the T arm. Among single-stranded nucleotides, A59 has a prominent role, but full expression of the gelonin-stimulating activity requires an extensive cooperation between nucleotides in both arms. |
| Databáze: | OpenAIRE |
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