Resumen:
We have described that acidic stress in pneumococcus induced
two different cellular processes. First, this pathogen induces an
acid tolerance response (ATR), and after triggers a programmed
cell death or autolysis. Recently, we also demonstrated the
involvement of F0.F1-ATPase in the ATR mechanism by
characterization of atp mutants (Cortes et al., poster presentation).
This enzyme facilitates the extrusion of protons from the cell
cytoplasm, preventing a drop in the intracellular pH in bacteria.
With the purpose to evaluate the F0.F1-ATPase effect on acidinduced
autolysis, wild-type cells were incubated at pH 7,6 with
10 μM DCCD, an specific inhibitor of this enzyme that blocks its
H+-translocating activity. We found that DCCD-induced autolysis
was similar to that one produced by acidic shock. To confirm the
role of this enzyme in the acid-induced autolysis, two F0.F1-ATPase
mutants were incubated at pH 5,6 and 37ºC for 6 hours. We
observed that the atpC G47V strain, which showed an increased
ATR, presented no lysis after 6 hours of incubation. On the contrary,
the atpC V48L strain, which had a decreased ATR, showed an
early induction of autolysis compared with the wild type strain.
According to these data we conclude that functionality of the F0.F1-
ATPase is required for preventing autolysis induced by acidic
stress. Work is in progress to study the checkpoints by which
pneumococcus, cultured under acidic conditions, pass from ATR
to autolysis, or omit ATR producing directly autolysis.