Open AccessMolecular Cloning and Expression of an Interferon‐Inducible Protein Encoded by Gene 203 from the Gene 200 Cluster
Author(s) -
Gribaudo Giorgio,
Ravaglia Stefania,
Guandalini Laura,
Riera Ludovica,
Gariglio Marisa,
Landolfo Santo
Publication year - 1997
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1997.t01-1-00258.x
Subject(s) - microbiology and biotechnology , untranslated region , complementary dna , polyadenylation , messenger rna , coding region , gene , biology , open reading frame , rapid amplification of cdna ends , interferon , gene expression , gene cluster , three prime untranslated region , molecular cloning , peptide sequence , genetics
We report here the complete coding sequence of a 203 cDNA, a member of the interferon‐inducible Ifi 200 gene family. By combining reverse‐transcriptase PCR and rapid amplification of cDNA ends (RACE) techniques we have obtained a 3.8‐kb cDNA corresponding to a 203 mRNA. When used as a probe in northern analysis, its 3’ segment hybridized to a 3.8‐kb interferon‐inducible mRNA, whereas the 5’‐end additionally hybridized to a less abundant interferon‐inducible 1.8‐kb mRNA. Nucleotide sequence analysis revealed that the two mRNAs share the 5’‐untranslated region and the same open reading frame, which encodes a hydrophilic protein composed of 408 amino acids. The difference between them is due to a 3’‐untranslated region extended by alternative polyadenylation site selection. Furthermore, 203 mRNA was found to be inducible by interferon‐α in various murine cell lines. Using polyclonal antibodies raised against a segment specific for the 203 protein, we established that p203 protein levels increase on treatment with interferon‐α in murine fibroblasts and that p203 is located in the nucleus.