Effect of Double-Stranded Viral RNA on Mammalian Cells in Culture

Abstract

During bovine enterovirus infection of Ehrlich ascites tumor cells, large amounts of doublestranded RNA accumulate. Addition of this doublestranded RNA to uninfected cells leads to rapid cell death. This is not a result of infectious virus production. Neither single-stranded RNA nor heat-denatured double-stranded RNA has this effect. Similar experiments with synthetic double-stranded polymers, poly(I) *poly(C) and poly(A). poly(U), show that they are only slightly toxic at the concentrations used. The effect of the double-stranded RNA is nonspecific for cells of different origins. The implications of this finding in relation to the cytopathic effects of picornavirus and to cancer chemotherapy are discussed.During the replication cycle of picornaviruses, doublestranded RNA is formed as an intermediate (1). In polio-and mengovirus infected cells, this double-stranded RNA is detectable 3-5 hr after infection (2, 3). In bovine enterovirus (BEV) replication, double-stranded RNA is detectable 2 hr after infection (manuscript in preparation). Unlike mengovirus-and poliovirus-infected cells, in which only 1-5% of the total amount of RNA is double-stranded, in BEV-infected cells, double-stranded RNA accumulates throughout the infection cycle; 7 hr after infection, just before cell death, it is the major species of viral RNA in the cells. In order to test whether there was any relationship between the accumulation of double-stranded RNA and the cytopathic effect, we have examined the effect of this RNA on various cells in culture. Our data suggest that viral double-stranded RNA is very toxic to such cells. MATERIALS AND METHODS Cells and virusMonolayer cultures of W138 were purchased from GIBCO (Grand Island, N.Y.) penicillin (100 units/ ml), streptomycin (100 gg/ml), and Fungizone (5 units/ml).Ehrlich ascites tumor and Sarcoma-1 were maintained as ascites tumors by intraperitoneal passage of about 108 cells at 8-day intervals in adult Swiss-Webster mice.Bovine enterovirus-1 (BEV) was grown as described (4). Titrations of the virus were performed on either L-cells or MDBK cells.

Poliovirus Temperature-sensitive Mutants Defective in Cytopathic Effects are also Defective in Synthesis of Double-stranded RNA

“…However, purified picornavirus DS-RNA rapidly reproduced the tb + character typical of picornavirus c.p.e. (Cordell-Stewart & Taylor, 1971). The kinetics were very close to those expected if DS-RNA itself were the cause of membrane damage, and heat denaturation (which liberates the cRNA strand) removed the cytotoxicity.…”

Action of spontaneously produced beta interferon in differentiation of embryonal carcinoma cells through an autoinduction mechanism.

“…dsRNA is known to be a potent biological effector (13). dsRNA can activate certain enzymes and induce transcription of some genes (26).…”

Double-Stranded RNA as an Inhibitor of Protein Synthesis and as a Substrate for a Nuclease in Extracts of Krebs II Ascites Cells

“…Addition of dsRNA at the same time as the mRNA inhibited protein synthesis directed by EMC RNA by about 50%, regardless of whether the S-30 itself was preincubated or not. When the S-30 was first incubated for 5 We have shown that dsRNA inhibits the translation of two natural mRNAs in an ascites cell-free system. This effect resembles that previously observed in reticulocyte lysates (6,7,12) in the following respects: (a) the rate of protein synthesis in the presence of dsRNA shows a sharp cutoff after an initial period of synthesis at the control rate (Fig.…”

“…Double-stranded RNA (dsRNA) is involved in several biological processes in animal cells. In particular, dsRNA induces the production of interferon (1-3), can cause tumor regression (4,5), and has been implicated in the shut-off of protein synthesis in cells infected with poliovirus (6,7). Several recent reports have demonstrated the binding and uptake of dsRNA by cells in tissue culture (8,9), and dsRNA has been found in normal rat liver (10) and in nuclei of sea urchin embryos (11).…”