Interferon messenger RNA content of human fibroblasts during induction, shutoff, and superinduction of interferon production.
Translation of injected mRNA in oocytes of Xenopus laevis has been used as a highly sensitive and quantitative assay for interferon mRNA. Injection into oocytes of polyadenylylated RNA extracted from poly(I).poly(C)-induced human diploid fibroblasts (FS-4) leads to the synthesis of biologically active human fibroblast interferon over a period of 24-32 hr. There is a linear relationship between the amount of mRNA injected and the interferon yield obtained over a range of 1-20 ng of injected RNA. Injection of 40-80 ng of mRNA into each of 15 oocytes, homogenized in 0.3 ml of incubation medium, gave a titer of 128-256 interferon reference units/ml of homogenate.FS-4 cells at the peak of interferon production-i.e., approximately 2.5 hr after the beginning of induction with poly(I).poly(C)-gave mRNA that yielded 24-48 interferon reference units/ml in the oocyte assay (30 ng of RNA injected per oocyte). An equivalent amount of mRNA from FS-4 cells in the shutoff phase, approximately 6 hr after induction, gave </=4 interferon reference units/ml. In contrast, mRNA extracted from FS-4 cells that had been induced and maintained in the presence of 40 muM 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole for 6 hr produced 64-128 interferon reference units/ml. Polyadenylylated RNA obtained from uninduced FS-4 cells did not lead to detectable interferon synthesis (<4 interferon reference units/ml). These data provide a direct verification of the hypothesis that the shutoff of interferon production in FS-4 cells involves a regulatory event leading to the posttranscriptional inactivation or degradation of interferon mRNA. Because the inactivating mechanism is sensitive to inhibition by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole, a selective inhibitor of nuclear heterogeneous RNA and mRNA synthesis, it is likely that synthesis of an RNA molecule is necessary for the shutoff of interferon production.
D000818:Animals; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007372:Interferons / Q000096:biosynthesis*; D007700:Kinetics; D007739:L Cells / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism*; D014981:Xenopus
D007093:Imidazoles; D012333:RNA, Messenger; D007372:Interferons; D003513:Cycloheximide
Science. 1969 Dec 19;166(3912):1474-80
17655040
Proc Natl Acad Sci U S A. 1964 Oct;52:1121-9
14224391
Virology. 1963 Nov;21:383-9
14081362
Science. 1959 Aug 21;130(3373):432-7
13675766
J Exp Med. 1954 Feb;99(2):167-82
13130792
Cell. 1976 Nov;9(3):473-80
1086720
Biochem Biophys Res Commun. 1977 Feb 21;74(4):1675-82
402922
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3287-91
269391
Proc Natl Acad Sci U S A. 1977 Apr;74(4):1483-7
266188
J Exp Med. 1977 Feb 1;145(2):344-56
833544
Virology. 1977 Jan;76(1):47-54
835243
Virology. 1976 Dec;75(2):384-93
1006987
Science. 1976 Oct 22;194(4263):431-3
982026
Nature. 1976 Mar 11;260(5547):141-3
176593
Nat New Biol. 1972 Sep 6;239(88):9-14
4404031
J Mol Biol. 1968 May 28;34(1):49-60
4106524
J Cell Biol. 1975 Dec;67(3):852-62
811672
Appl Microbiol. 1971 Apr;21(4):723-5
4325022
J Cell Biol. 1976 May;69(2):229-40
1262389
Proc Natl Acad Sci U S A. 1975 Jun;72(6):2185-7
49055
J Biol Chem. 1976 Mar 25;251(6):1659-62
176158
J Exp Med. 1975 Nov 1;142(5):1283-1300
53261
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3909-13
4544057
J Mol Biol. 1972 Oct 28;71(1):21-30
4539023
Proc Natl Acad Sci U S A. 1975 Oct;72(10):4157-61
172898
Proc Natl Acad Sci U S A. 1975 Oct;72(10):3975-7
172890
Cell. 1974 Nov;3(3):255-64
4434473
J Biol Chem. 1973 Mar 10;248(5):1502-12
4694721
Antimicrob Agents Chemother. 1972 Dec;2(6):476-84
4670440
J Cell Biol. 1975 Dec;67(3):835-51
811671
Proc Natl Acad Sci U S A. 1976 May;73(5):1621-5
1064033
Proc Natl Acad Sci U S A. 1976 May;73(5):1442-6
1064017
J Infect Dis. 1976 Jun;133 Suppl:A22-9
778308
Proc Natl Acad Sci U S A. 1975 Dec;72(12):4881-5
1061077
Proc Natl Acad Sci U S A. 1975 Nov;72(11):4285-7
1060107
Proc Natl Acad Sci U S A. 1975 Oct;72(10):3898-901
1060071
Proc Natl Acad Sci U S A. 1976 Feb;73(2):520-523
1061154
Proc Natl Acad Sci U S A. 1976 Jan;73(1):29-33
1061124
Virology. 1976 Apr;70(2):542-4
1266051
Virology. 1976 Apr;70(2):532-41
1266050
Virology. 1976 Mar;70(1):256-9
1258381
Annu Rev Microbiol. 1975;29:131-61
1180510
Science. 1975 Oct 17;190(4211):282-4
1179208
Virology. 1975 Feb;63(2):475-83
1168006
Proc Natl Acad Sci U S A. 1970 Sep;67(1):464-71
5272327
Proc Natl Acad Sci U S A. 1967 Sep;58(3):1004-10
5233831
Nature. 1969 May 17;222(5194):682-3
5768287