Simian virus 40 and polyoma virus induce synthesis of heat shock proteins in permissive cells.

Abstract

During the lytic infection of monkey and mouse cells with simian virus 40 and polyoma virus, respectively, the preferentially increased synthesis of two host proteins of 92,000 and 72,000 Mr was observed by 15 to 20 h after infection besides the general stimulation of most cellular proteins. In a variety of organisms and cell cultures, from bacteria and yeasts to humans, a mild heat shock was found to induce the vigorous synthesis of a few characteristic proteins. In Drosophila melanogaster, where this phenomenon has been extensively investigated, the induction of the heat shock proteins is paralleled by a strong reduction in the synthesis of most proteins made before the heat shock. In avian and mammalian cells, decreased protein synthesis is less evident, but the increased synthesis of at least two proteins in the range of 85,000 to 95,000 Mr and 70,000 to 75,000 Mr was observed; these proteins may be related to two of the major heat shock proteins in insects. The increased synthesis of the same or similar proteins has been reported for cells subjected to a variety of treatments, such as exposure to amino acid analogs, chelating drugs, heavy metal ions, arsenite, and other sulfhydryl reagents (for a review, see reference 19).The lytic infection of mouse and monkey cell cultures with polyoma virus or simian virus 40 (SV40), respectively, induces an increased synthesis of the majority of cellular proteins (9). Among these, two strongly stimulated host proteins fall into the same size range as the proteins mentioned above (E. W. Khandjian, P. Arrigo, T. Rose, and J.-M. Matter, Experientia 36:749, 1980). Here, we show that the two proteins that are stimulated by virus infection are also induced by thermal treatment of uninfected mouse and monkey cells. MATERIALS AND METHODSCell cultures, virus infections, and thermal treatment. Confluent primary mouse kidney cultures and confluent cultures of CV-1 (African green monkey) cells in 9-cm-diameter petri dishes were mock infected or infected with polyoma virus or SV40, respectively, and incubated at 37°C as described previously (9). For thermal treatment (heat shock), cultures were incubated at 43.5°C for 1 h before labeling at 37°C for 2 h with 100 iCi of [35S]methionine (500 to 1,000 Ci/mmol; The Radiochemical Centre, Amersham, United Kingdom) in 2 ml of methionine-free medium.Extraction of proteins. After [35S]methionine labeling, the cell monolayers were washed with cold phosphate-buffered saline, total proteins were solubilized by the addition of sodium dodecyl sulfate (SDS) sample buffer (68 mM Tris-hydrochloride [pH 6.8], 2% SDS, 2% 2-mercaptoethanol, 0.01% bromophenol blue, 15% glycerol) (10), and the viscous extract was sonicated. Alternatively, cultures put on ice were lysed with 0.5% Nonidet P-40 in 0.1 M Tris-hydrochloride (pH 9)-0.1 M NaCI-5 mM MgCI2 for 10 min, and the lysates were centrifuged at 30,000 x g for 30 min at 4°C. To the supernatant 0.5 volume of threefoldconcentrated SDS sample buffer was added, and the samples were heated in a boiling water ...

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).…”

Enhancement of the Interferon‐Induced Double‐Stranded RNA‐Dependent Protein Kinase Activity by Sindbis Virus Infection and Heat‐Shock Stress

“…Cultured cells sometimes respond to viral infection with stress responses, such as synthesis of heat-shock proteins (2,8,9,11,17,18,20). The activation of PK-1 in the cells infected with Sindbis virus may then be one of such stress responses.…”

Rapid purification of mammalian 70,000-dalton stress proteins: affinity of the proteins for nucleotides.

“…Hence, it has been suggested by us and others that the stress proteins most likely serve functions in the cell which are distinct from their participation during stress. Indeed, synthesis and post-translational modification of some of the stress proteins have been shown to be sensitive to: (i) extracellular levels of either glucose or calcium (25); (ii) viral infection (7,11,15); and (iii) various growth factors or mitogens (22 Much of the work examining the structure and function of the stress proteins has focused on the ca. 70-kDa proteins.…”

“…Indeed, synthesis and post-translational modification of some of the stress proteins have been shown to be sensitive to: (i) extracellular levels of either glucose or calcium (25); (ii) viral infection (7,11,15); and (iii) various growth factors or mitogens (22).…”