Full‐length sequence and expression of the 42 kDa 2–5A synthetase induced by human interferon

FEBS Letters

Cravador, A.¹  DeWit, L.²  Content, J.²  Wathelet, M.¹  Defilippi, P.²  Moutschen, S.²  Huez, G.¹ 

[1] Département de Biologie Moléculaire, Université Libre de Bruxelles, 67 rue des Chevaux, B-1640 Rhode-St-Genèse, Bruxelles, Belgium;Département de Virologie, Institut Pasteur du Brabant, 642 rue Engeland, B-1180 Bruxelles, Belgium

关键词: Interferon induction;    cDNA;    Cloning;    Transcription;    Translation;   

DOI  :  10.1016/0014-5793(86)80224-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown