Full-length sequence and expression of the 42 kDa 2-5A synthetase induced by human interferon
| Autor: | M. Wathelet, S. Moutschen, A. Cravador, L. DeWit, P. Defilippi, G. Huez, J. Content |
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| Rok vydání: | 1986 |
| Předmět: |
Translation
Cytoplasm Microinjections RNA Splicing DNA Recombinant Biophysics Biology Biochemistry Xenopus laevis Structural Biology Transcription (biology) Complementary DNA 2' 5'-Oligoadenylate Synthetase Genetics Animals Humans Coding region Amino Acid Sequence RNA Messenger Molecular Biology Gene Cellular localization Cell Nucleus Nucleoplasm Base Sequence Cell-Free System RNA DNA Cell Biology Molecular biology Gene Expression Regulation Enzyme Induction Interferon Type I RNA splicing Oocytes Female Interferon induction cDNA Transcription Cloning |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/0014-5793(86)80224-1 |
| Popis: | Interferon-induced 2–5A synthetases are probably involved in some antiviral actions of interferon. In human cells two different mRNAs (1.6, 1.8 kb long) coding for this protein are transcribed from the same gene and are produced by differential splicing. The relationship between the two mRNAs of different size and the active enzyme is not clear, nor is the cellular localization of the latter known. We have cloned a cDNA corresponding to the 1.6 kb RNA. This cDNA was sequenced and its complete coding region was subcloned into pSP64. The resulting plasmid was used to direct the synthesis of micrograms of capped RNA transcript after linearization in the 3'-non-coding region. A 39 kDa protein was synthesized when this RNA was translated in rabbit reticulocyte lysate. When this capped RNA was introduced by microinjection into Xenopus oocytes, production of 2–5A synthetase was clearly observed in the cytoplasm and 10–30% of the enzyme accumulated with time in the nucleoplasm. Analysis of cytoplasmic homogenates of these oocytes on a glycerol gradient revealed that the enzyme is fully active in the monomeric form. |
| Databáze: | OpenAIRE |
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