Our results support a model in which c-KIT signaling is targeted by Yersinia T3SS to suppress pro-inflammatory
responses. Some kinases activated downstream of c-KIT, such as MEK and PI3K, have been shown to be inhibited by the Yersinia effectors YopJ and YopH, respectively [9, 10, 42]. YopJ has also been shown to inhibit phosphorylation of MKK4/SEK1 and attenuates JNK signaling and subsequent Regorafenib EGR1 activation  (Figure 8). Our findings suggest that downregulation of a receptor kinase function that leads to NF-κB activation can ameliorate the inhibitory effect of Yersinia T3SS. Since we observed that the inhibition of another signaling protein AKT1 also resulted in higher production of TNF-α by Yersinia-infected macrophage cells (Figure 3), we hypothesized that upon bacterial infection, multiple signal transduction pathways are triggered by various host extracellular and intracellular receptors of pathogen associated selleck chemicals molecular patterns (PAMPs). However, not all signaling pathways are inactivated by Yersinia during infection, and inhibition of c-KIT may lead to redirection to alternative signaling pathways, such as the LPS-activated
CD14 and TLR4 signaling to p38 and JNK, to recover selleckchem NF-KB-driven gene expression [44, 45]. This hypothesis is supported by our observations that pharmacological inactivation of JNK1 using the inhibitor BI-78D3 did not recover pro-inflammatory gene expression in THP-1 cells infected with pathogenic Yersinia (Figure 5A), while AKT1 and c-KIT inhibition resulted in increased TNF-α production in infected THP-1 and NHDC (Figure 3). Thus, redistribution of signaling pathways can still lead to mitigation of NF-κB-regulated immune response during the course of Yersinia infection. The exact mechanism of Yersinia activation of c-KIT remains unclear. The natural ligand of c-KIT, SCF, has been shown to activate c-KIT phosphorylation within 5 min of treatment [34, 35]. In response to Y. enterocolitica, c-KIT exhibited maximal phosphorylation at ~45 min post-infection in THP-1 cells by Western blot (Figure 6), demonstrating that Yersinia infection is capable of stimulating c-KIT activation,
albeit via a delayed response compared to SCF. Since, we observed this delayed phosphorylation in both virulent Fenbendazole and attenuated Y. enterocolitica, it may be the case that LPS or other bacterial cell surface molecule can mediate host receptor phosphorylation and/or signaling, rather than solely the T3SS. We have also shown that inhibition of c-KIT signaling by the small molecule OSI-930 induced an altered inflammatory gene expression pattern in response to pathogenic Yersinia that resembled infection by a non-virulent strain (Figure 5A), further supporting functional links between c-KIT activity and Yersinia virulence. It may be the case that Yop effectors either directly or indirectly modulate c-KIT function following injection into the host.