The protein pattern of a type-D retrovirus (PMFV) isolated from and propagated in human cell lines has been investigated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Staining with Coomassie blue and labeling with 14C-leucine/14C-lysine revealed five viral polypeptides with molecular weights of 10,000, 12,000, 15,000, 25,000, and 68,000. The 68,000 D-protein was shown to be a glycoprotein by incorporation of 3H-glucosamine and the 15,000 D-protein was identified as a phosphoprotein. By comparing PMFV with the closely related Mason-Pfizer monkey virus (MPMV) in co-electrophoresis experiments no clear difference was detected in viral 14C-leucine/14C-lysine profiles. The viruses differ, however, with respect to their glycoprotein patterns. A glycoprotein corresponding to the gp20 of MPMV has not been detected in PMFV irrespective of the cell line used for propagation of viruses.
Exposure in vitro of various mammalian retroviruses to the chelating agents EDTA or EGTA in millimolar concentrations resulted in partial disintegration of viral membranes as measured by accessibility or even release of reverse transcriptase, an internal viral protein, without any other treatment usually required. Among the viruses responding to chelators were mammalian type C viruses, primate type D viruses and bovine leukemia virus. The effect was dose-dependent. The avian type C virus AMV, however, was found to be not susceptible to the agents. Rauscher mouse leukemia virus treated in vitro with EDTA or EGTA showed reduced infectivity in mice. The results are considered as evidence for some association of divalent cations with membranes of mammalian retroviruses. The disintegrating activity of EGTA suggests that Ca2+ is an integral constituent of viruses but Mg2+ may also be involved. These cations seem to be responsible for maintaining integrity of retroviral membranes which, after chelation of ions, are either disrupted or become permeable for the exogenous template of reverse transcriptase. In addition, the disintegrating activity of trifluoperazine may indicate that a calmodulin-like protein occurs in retroviral membranes.
Normal and precancerous livers as well as primary and transplanted hepatomas are studied for glycogen content, hexokinase- and glucokinase activity, at the same time measuring glycolysis in the high-speed supernatants of these tissues with different substrates and additives.Experimental results show that:1) normal and precancerous rat livers have low hexokinase activity and low glycolysis, but high glucokinase activity and high glycogen content;2) primary DAB hepatomas. on the contrary, have high hexokinase activity and raised glycolysis as well as low glycokinase activity and low glycogen content;3) transplantable DAB- as well as DENA-hepatomas have high hexokinase activity and high glycolyses. while neither glucokinase activity nor glycogen content have been found;4) when using glucose as substrate and adding exogenous hexokinase. there is a considerable increase in formation of lactic acid in all tissue supernatants investigated;5) if glucose-6-phosphate is used instead of glucose, a considerable increase of glycolysis is noted as well.It is concluded from the results obtained that the hexokinase reaction limits the extent of glycolysis in these tissues.
In contrast to the human cell line derived type D retrovirus PMFV, the Mason-Pfizer monkey virus (MPMV) does not suppress the mitogen response of normal human lymphocytes. Both viruses have been propagated on the same cell lines and purified by the same methods. MPMV did not contain a factor able to abolish PMFV-induced suppression of the mitogen response. Neither could MPMV suppress the mitogen response of lymphocytes from rhesus monkeys or baboons. PMFV however inhibited their reactivity.
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