Open AccessCorrelation between the Induction of Mouse Interferon and the Amount of Its mRNA
Author(s) -
RAJ N. Babu Kishan,
FERNIE Bruce F.,
PITHA Paula M.
Publication year - 1979
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1979.tb13179.x
Subject(s) - interferon , cycloheximide , microbiology and biotechnology , messenger rna , biology , rna , fibroblast , chemistry , protein biosynthesis , biochemistry , virology , gene , in vitro
Injection into Xenopus oocytes of polyadenylated cytoplasmic RNA from mouse C‐243‐3 cells induced with poly(rI) · poly(rC) and DEAE‐dextran resulted in the production of biologically active interferon. The synthesis of interferon by oocytes is linearly related to the amount of mRNA injected. Thus a titre of 160 reference units/oocyte was obtained by injecting 250 ng of total cytoplasmic poly(A)‐containing RNA. The kinetics of interferon production in mouse C‐243‐3 cells induced with poly(rI) · poly(rC) and DEAE‐dextran are characterized by an initial lag of 3–4 h and a burst of interferon secretion between 5–8 h with a maximum accumulation at 7 h. Comparison of the relative amount of interferon mRNA at various time intervals of induction with the cellular synthesis of interferon showed a general correlation. At 9 h after induction, i.e. during the shut off phase of interferon production, the cells contained no detectable amount of interferon mRNA (< 20 units/ml). Attempts to superinduce interferon production in mouse C‐243‐3 cells by using the cycloheximide/actinomycin D protocol as in human fibroblast cells were not successful. α‐Amanitin at concentrations similar to those required to inhibit cellular DNA‐dependent RNA polymerase form II completely inhibited the interferon production when applied during the first 3 h of induction. Addition of α‐amanitin at 3, 5 or 7 h after induction stimulated the subsequent interferon production by 2–4‐fold.