3 research outputs found
Functional analysis of the SseK family of Salmonella SPIâ2 type III secretion system effectors
In humans, serovars of Salmonella enterica cause diseases ranging from gastroenteritis to
systemic typhoid fever. Salmonella encodes numerous translocated virulence (effector) proteins
that enable the bacteriumâs complex intracellular lifestyle. Three poorly characterised Salmonella
effectors are the SseK proteins: SseK1, SseK2 and SseK3. This work showed that all three effectors
are translocated by the SPIâ2 type III secretion system into macrophages. Despite high amino
acid similarity between the proteins, SseK1 is cytosolic and SseK2 and SseK3 associate with the
Golgi network. When ectopically expressed in mammalian cells, SseK1, SseK2 and SseK3
suppressed NFâÎșB pathway activation. This inhibition was specific for TNFαâinduced signaling
but not TLR4â, or interleukinâinduced NFâÎșB pathway activation. After physiological delivery
of the effectors into macrophages during Salmonella infection, SseK1 and SseK3 inhibited NFâ
ÎșB activity in an additive manner and suppressed TNFαâinduced necroptotic host cell death.
Despite stable interaction between SseK3 and the E3âubiquitin ligase TRIM32 in macrophages,
SseK3âmediated inhibition of NFâÎșB activation and host cell death did not require TRIM32. In
addition, bacterial burden in Trim32â/â mice were indistinguishable from wildâtype mice after
shortâterm infection with Salmonella. Catalytic activity of the SseK effectors required a DxD
motif, which in SseK1 and SseK3 is essential for arginineâNâacetylglucosamine (GlcNAc)â
ylation of target proteins. In infected macrophages, SseK1 and SseK3 caused arginineâ
GlcNAcylation of a diverse range of distinct bacterial and host proteins, including autoâ
GlcNAcylation of SseK1 and SseK3. Both SseK1 and SseK3 induced arginineâGlcNAcylation of
TRADD, which is most likely the cause of the effectorâs inhibitory effect on both NFâÎșB
activation and host cell death. In addition, a large number of Golgi network associated
proteins were identified as potential SseK3 GlcNAcylation targets. This suggests additional,
yetâtoâbe validated functions of the effectors during Salmonella infection besides inhibition
of NFâÎșB signalling and necroptosis in macrophages.Open Acces