Infection with paramyxoviruses stimulates synthesis of cellular polypeptides that are also stimulated in cells transformed by Rous sarcoma virus or deprived of glucose

Abstract

ABSTACInfection of several types of cultured cells with the paramyxoviruses simian virus 5 and Sendai virus stimulates synthesis of four polypeptides (I-IV) with molecular weights of approximately 99,000,97,000, 86,000, and 78,000, respectively. That these are host polypeptides encoded in cellular mRNAs has been shown by the inhibition of their synthesis by actinomycin D and by the similarity of the peptide maps of them and of polypeptides with the same electrophoretic mobility from uninfected cells. Peptide mapping as well as identical migration in polyacrylamide gels has also indicated that polypeptides I, II, and IV are the same as plasma membrane polypeptides whose synthesis is enhanced in cells transformed by Rous sarcoma virus and in normal cells by glucose deprivation or treatment with 2-deoxyglucose. Polypeptides I and II appear to be the same polypeptides, with the observed differences in migration reflecting the glycosylation of polypeptide I, a relationship-previously shown to exist between polypeptides in glucose-deprived and glucose-fed cells. Infection with paramyxoviruses does not significantly increase the transport of glucose by cells, and the maintenance of a high concentration of glucose in the medium does not prevent the enhanced synthesis of these polypeptides. This is in contrast to the situation in transformedcells in which stimulation of synthesis of these polypeptides is secondary to depletion of glucose in the mediuwr due to increased glucose uptake by the cells. Thus, although paramyxovirus infection and transformation by Rous sarcoma virus result in stimulation of the synthesis of the same membrane polypeptides, the mechanism of stimulation differs.We have reported (1) that, in addition to the viral structural polypeptides and a nonstructural polypeptide, four polypeptides were synthesized in increased amounts in cells infected with the paramyxovirus simian virus 5 (SV5). These polypeptides, designated I-IV, had estimated molecular weights of 99,000, 97,000, 86,000, and 78,000, respectively. It was suggested that these were cellular polypeptides whose synthesis was enhanced by infection because: (i) (4, 7). On the basis of these results, these polypeptides have been referred to as "glucose-regulated proteins" (4). As described here, paramyxovirus infection, unlike transformation, does not cause a significant increase in glucose transport, and maintenance of a high glucose concentration does not prevent the enhanced synthesis of these polypeptides in paramyxovirusinfected cells. Thus, paramyxovirus infection apparently produces a perturbation in the cell membrane, different from transformation, that alters the rate of synthesis of these polypeptides.MATERIALS AND METHODS Cells. CV-1 cells (TC7 clone) were grown in Eagle's medium with 10% fetal calf serum, and HeLa cells were grown in reinforced Eagle's medium with 5% fetal calf serum as described (1) (Eagle's medium contains 5.5 mM glucose and the reinforced medium contains 22 mM glucose). Primary chicken embryo fibroblasts (CEFs) a...

Abnormal Forms of the Herpes Simplex Virus Immediate Early Polypeptide Vmw175 Induce the Cellular Stress Response

“…The induction of the stress response by viruses other than HSV also appears to involve a different mechanism, since wt viruses mediate the effect. This is true of the DNA-containing adenoviruses and papovaviruses (Nevins, 1982;Khandjian & Turler, 1983), as well as RNA-containing viruses (Hightower & Smith, 1978;Peluso et al, 1978 ;Collins & Hightower, 1982;Garry et al, 1983). In the case of adenoviruses, the E1A gene product is thought to activate stress protein gene expression by interaction with DNA sequences in the promoter (Kao & Nevins, 1983;Kingston et al, 1984).…”

“…The stimuli include infection with bacteriophage or other viruses (Hightower & Smith, 1978;Peluso et al, 1978;Collins & Hightower, 1982;Drahos & Hendrix, 1982;Nevins, 1982;Notarianni & Preston, 1982;Garry et al, 1983;Khandjian & Turler, 1983;LaThangue et al, 1984). Although the precise function of the stress response or the stress proteins themselves is unclear, it is thought that the stress response has a protective role, since stressed cells can more readily withstand a subsequent exposure to the agents which induced the response (Li & Werb, 1982;Schlesinger, 1986).…”

Analysis in vivo of GRP78-BiP/substrate interactions and their role in induction of the GRP78-BiP gene.

“…The polypeptides identified by these groups were later shown to be identical, thus linking its glucose-regulated expression with a biochemical property (Munro and Pelham, 1986). Other cellular perturbations that result in an induction of GRP78-BiP synthesis include inhibition of N-linked glycosylation and modification Watowich and Morimoto, 1988), disruption of intracellular calcium stores (Drummond et al, 1987), infection by enveloped viruses (Peluso et al, 1978;Stoeckle et al, 1988), the expression in the ER of mutant polypeptides deficient in folding (Kozutsumi et al, 1988), and the increased expression of secretory proteins (Dorner et al, 1989;Wiest et al, 1990). These treatments affect processes in the ER where GRP78-BiP is localized, which suggests the existence of a feedback loop to regulate the expression of the gene.…”

Effect of Vesicular Stomatitis Virus and Semliki Forest Virus on Uptake of Nutrients and Intracellular Cation Concentration

“…The infection of cells by cytolytic, nontransforming viruses such as vesicular stomatitis virus (VSV) or Semliki Forest virus (SFV) might be thought to have, if anything, an opposite effect on membrane function in that the synthesis of host DNA, RNA and protein becomes depressed (Pfefferkorn & Shapiro, 1974;Wagner, 1975), rather than elevated (but see Peluso et al, 1977). However, we show here that BHK cells infected with VSV or SFV accumulate 2-deoxy-D-glucose two to three times more effectively than uninfected cells.…”