Interferon-gamma receptor: mRNA half-life, receptor mass, and abundance on A431 human epidermoid carcinoma cells.
The human interferon-gamma (IFN-gamma) receptor protein and mRNA were identified in the epidermoid carcinoma cell line A431 by radioligand binding and Northern hybridization, respectively. Receptor affinity and receptor number per cell were calculated by Scatchard analysis of saturation binding data. The half-life of the receptor mRNA was determined by actinomycin D inhibition of de novo transcription. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of covalently cross-linked 32P-labeled IFN-gamma and its receptor is consistent with a molecular mass of approximately 99 kD. Radioligand binding analysis revealed 27,000 high affinity (KD = 7.1 x 10(-10) M) receptors per cell. Northern hybridization using an IFN-gamma receptor cDNA probe revealed a single mRNA of 2.2 kb. The receptor mRNA half-life in actinomycin D-treated cells was 4.7 h.
Crouse
J A
JA
Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN 37232.
Mitchell
W M
WM
D015152:Blotting, Northern; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000378:metabolism; D003609:Dactinomycin; D004591:Electrophoresis, Polyacrylamide Gel; D006207:Half-Life; D006801:Humans; D007372:Interferons; D008970:Molecular Weight; D012333:RNA, Messenger / Q000378:metabolism*; D011869:Radioligand Assay; D011971:Receptors, Immunologic / Q000032:analysis / Q000235:genetics*; D017471:Receptors, Interferon; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D003609:Dactinomycin; D007372:Interferons