title: Isolation and characterization of a new mutant human cell line unresponsive to alpha and beta interferons.
abstract: Previously we described human cell line 2fTGH, in which expression of guanine phosphoribosyltransferase is tightly controlled by the upstream region of interferon (IFN)-stimulated human gene 6-16. After mutagenesis of 2fTGH and selection with 6-thioguanine and IFN-alpha, we isolated 11.1, a recessive mutant that does not respond to IFN-alpha. We now describe U2, a second recessive mutant, selected similarly, that complements 11.1. U2 had no response to IFN-alpha or IFN-beta, and its response to IFN-gamma was partially defective. Although many genes did respond to IFN-gamma in U2, the 9-27 gene did not and the antiviral response of U2 cells to IFN-gamma was greatly reduced. Band shift assays showed that none of the transcription factors normally induced in 2fTGH cells by IFN-alpha (E and M) or IFN-gamma (G) were induced in U2. However, extracts of untreated U2 cells gave rise to a novel band that was increased by treatment with IFN-gamma but not IFN-alpha. Band shift complementation assays revealed that untreated and IFN-gamma-treated U2 cells lack the functional E gamma subunit of transcription factor E and that IFN-alpha-treated U2 cells do contain the functional E alpha subunit.
authors.affiliation: Imperial Cancer Research Fund, London, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: John
authors.forename: R
authors.initials: R
authors.lastname: McKendry
authors.forename: S
authors.initials: S
authors.lastname: Pellegrini
authors.forename: D
authors.initials: D
authors.lastname: Flavell
authors.forename: I M
authors.initials: IM
authors.lastname: Kerr
authors.forename: G R
authors.initials: GR
authors.lastname: Stark
mesh_terms: D000585:Aminacrine / Q000031:analogs & derivatives / Q000494:pharmacology; D002459:Cell Fusion; D002460:Cell Line; D005808:Genes, Recessive; D005816:Genetic Complementation Test; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology; D016296:Mutagenesis; D009153:Mutagens / Q000494:pharmacology; D009588:Nitrogen Mustard Compounds / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013866:Thioguanine / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D007370:Interferon Type I; D009153:Mutagens; D009588:Nitrogen Mustard Compounds; D012333:RNA, Messenger; D000585:Aminacrine; D007371:Interferon-gamma; C004294:acridine half-mustard; D013866:Thioguanine