A mutant cell line partially responsive to both IFN-alpha and IFN-gamma.
A recessive mutant cell line, B7, which is partially responsive to both interferon (IFN)-alpha and IFN-gamma is described. B7 was FACS sorted from a cellular pool, which was obtained from the parental cell line 2C4, after several rounds of mutagenesis. The partial responsiveness to IFN was observed both in terms of expression of cell surface markers (CD2, class I and II HLAs) and mRNA expression of IFN-stimulated genes (9-27; 6-16; 2'-5' OAS; GBP and HLA-DR alpha). A genetic cross with the U4 mutant (JAK1-, a member of the Janus family of nonreceptor tyrosine kinase) did not restore full IFN responsiveness to B7, and JAK1 cDNA transfection into B7 restored the wild phenotype of the cell line, defining B7 as a member of the U4 complementation group. Nevertheless, JAK1 mRNA was not detected in this mutant. Transcriptional regulator complexes such as IRF1/2 (IFN-regulatory factor) and ISGF3-gamma (IFN-stimulated gene factor) were constitutively formed in the B7 mutant and co-migrated with the IFN-induced complexes expressed in the parental cell line 2C4. Thus, this cell line seems to be useful for understanding cis-acting elements governing JAK1 mRNA expression.
Bonjardim
C A
CA
Departamento de Microbiologia, Universidade Federal de Minas Gerais, Brasil. claubonj@mono.icb.ufmg.br
D018929:Cell Culture Techniques; D002461:Cell Line, Transformed / Q000187:drug effects*; D018076:DNA, Complementary / Q000302:isolation & purification; D004586:Electrophoresis; D005434:Flow Cytometry; D016898:Interferon-alpha / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology*; D009154:Mutation; D015533:Transcriptional Activation / Q000187:drug effects
D018076:DNA, Complementary; D016898:Interferon-alpha; D007371:Interferon-gamma