INVESTIGADORES
SARNACKI Sebastian Hernan
congresos y reuniones científicas
Título:
Impaired expression and secretion of sopB, sopD and sopE2 in Salmonella dam mutants
Autor/es:
GIACOMODONATO MN; GARTNER AS; GARCÍA BLATZ MD; SARNACKI SH; NOTO LLANA M; CERQUETTI MC
Lugar:
Portero de Funes, San Luis
Reunión:
Congreso; XLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2011
Institución organizadora:
Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB)
Resumen:
Dam methylation is an essential factor involved in the
virulence of an increasing number of bacterial pathogens including Salmonella
spp. dam mutants have a reduced capacity to colonize murine organs
compared to the parental strain. This partial inability could be related to a
reduced secretion of invasion proteins. Then, we investigated the involvement
of Dam methylation on the expression and secretion of essential proteins for
invasion, such as SopB, SopD and SopE2. These effectors are located outside of
SPI-1 but are translocated in a SPI-1-dependent manner. S.
Typhimurium sopB, sopD and sopE2 tagged (3xFLAG)
strains and their derivative Δdam were used during the experiments. The
expression and secretion of these effectors were determined in vitro, under
SPI-1 culture conditions, analyzing bacterial pellets and culture supernatants,
respectively. Tagged proteins were examined by SDS-page and immunoblotting with
anti-FLAG antibodies. Western blot results showed an impaired SopB, SopD and
SopE2 expression and secretion in dam mutants with respect to the
parental strain (p < 0.05). These results were further confirmed by QRT-PCR
showing that sopB, sopD and sopE2 genes transcription were two or
three-fold lower in dam mutants compared to wild type strain. Taken
together, our results demonstrate that Dam methylation modulates the expression
and secretion of SopB, SopD and SopE2.