Inactivation of the ptsI gene encoding enzyme I of the sugar phosphotransferase system of Streptococcus salivarius: effects on growth and urease expression
Ca. Weaver et al., Inactivation of the ptsI gene encoding enzyme I of the sugar phosphotransferase system of Streptococcus salivarius: effects on growth and urease expression, MICROBIO-UK, 146, 2000, pp. 1179-1185
The urease genes of Streptococcus salivarius 57.1 are tightly repressed in
cells growing at neutral ph. When cells are cultivated at acidic ph values,
the urease genes become derepressed and transcription is enhanced when cel
ls are growing under carbohydrate-excess conditions. Previously, the author
s proposed that the bacterial sugar: phosphotransferase system (PTS) modula
ted the DNA-binding activity by phosphorylation of the urease repressor whe
n carbohydrate was limiting. The purpose of this study was to assess whethe
r enzyme I (EI) of the PTS could be involved in modulating urease expressio
n in response to carbohydrate availability. An El-deficient strain (ptsl18-
3) of S. salivarius 57.1 was constructed by insertional inactivation of the
ptsl gene. The mutant had no measurable PTS activity and lacked El, as ass
essed by Western analysis. The mutant grew as well as the wild-type strain
on the non-PTS sugar lactose, and grew better than the parent when another
non-PTS sugar, galactose, was the sole carbohydrate. The mutant was able to
grow with glucose as the sole carbohydrate, but displayed a 24 h lag time
and had a generation time some threefold longer than strain 57.1. The mean
OD600 attained after 48 h by ptsl18-3 supplied with fructose was 0.16, with
no additional growth observed even after 3 d. Urease expression in the wil
d-type and mutant strains was assessed in continuous chemostat culture. Rep
ression of urease at neutral ph was seen in both strains under all conditio
ns tested. Growth of wild-type cells on limiting concentrations of lactose
resulted in very low levels of urease expression compared with growth on PT
S sugars. In contrast, under similar conditions, urease expression in ptsl1
8-3 was restored to levels seen in the parent growing on PTS sugars. Growth
under conditions of lactose excess resulted in further derepression of ure
ase, but ptsl18-3 expressed about threefold higher urease activity than 57.
1. The results support a role for El in urease regulation, but also indicat
e that additional factors may be important in regulating urease gene expres
sion.