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(Received for publication, August 7, 1996, and in revised form, October 3, 1996)
From the Madin-Darby canine kidney cells infected with
Sendai virus rapidly lose GSH without increase in the oxidized
products. The reduced tripeptide was quantitatively recovered in the
culture medium of the cells. Since the GSH loss in infected cells was not blocked by methionine, a known inhibitor of hepatocyte GSH transport, a nonspecific leakage through the plasma membrane is proposed. UV-irradiated Sendai virus gave the same results, confirming that the major loss of GSH was due to membrane perturbation upon virus
fusion. Consequent to the loss of the tripeptide, an intracellular pH
decrease occurred, which was due to a reversible impairment of the
Na+/H+ antiporter, the main system responsible
for maintaining unaltered pHi in those cells. At the end of
the infection period, a rise in both pHi value and GSH
content was observed, with a complete recovery in the activity of the
antiporter. However, a secondary set up of oxidative stress was
observed after 24 h from infection, which is the time necessary
for virus budding from cells. In this case, the GSH decrease was partly
due to preferential incorporation of the cysteine residue in the viral
proteins and partly engaged in mixed disulfides with intracellular
proteins. In conclusion, under our conditions of viral infection,
oxidative stress is imposed by GSH depletion, occurring in two steps
and following direct virus challenge of the cell membrane without the
intervention of reactive oxygen species. These results provide a
rationale for the reported, and often contradictory, mutual effects of
GSH and viral infection.
Volume 272, Number 5,
Issue of January 31, 1997
pp. 2700-2708
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
,
Institute of Biochemical Sciences,
University of Chieti "G. D'Annunzio," 66100 Chieti, Italy, the
§ Department of Experimental Medicine and Biochemical
Sciences, University of Rome "Tor Vergata," 00133 Rome, Italy,
and the ¶ Department of Biology, University of Rome "Tor
Vergata," 00133 Rome, Italy
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