Journal Article

Inflammasome Activation in Response to the Yersinia Type III Secretion System Requires Hyperinjection of Translocon Proteins YopB and YopD

Snyder, Annelise G., Wynosky-Dolfi, Meghan A., Philip, Naomi H., Zwack, Erin E., Ruthel, Gordon, Bliska, James B., Marketon, Melanie M., Francis, Matthew S., Brodsky, Igor E.

2015

Overview

Type III secretion systems (T3SS) translocate effector proteins into target cells in order to disrupt or modulate host cell signaling pathways and establish replicative niches. However, recognition of T3SS activity by cytosolic pattern recognition receptors (PRRs) of the nucleotide-binding domain leucine rich repeat (NLR) family, either through detection of translocated products or membrane disruption, induces assembly of multiprotein complexes known as inflammasomes. Macrophages infected with Yersinia pseudotuberculosis strains lacking all known effectors or lacking the translocation regulator YopK induce rapid activation of both the canonical NLRP3 and noncanonical caspase-11 inflammasomes. While this inflammasome activation requires a functional T3SS, the precise signal that triggers inflammasome activation in response to Yersinia T3SS activity remains unclear. Effectorless strains of Yersinia as well as Δ yopK strains translocate elevated levels of T3SS substrates into infected cells. To dissect the contribution of pore formation and translocation to inflammasome activation, we took advantage of variants of YopD and LcrH that separate these functions of the T3SS. Notably, YopD variants that abrogated translocation but not pore-forming activity failed to induce inflammasome activation. Furthermore, analysis of individual infected cells revealed that inflammasome activation at the single-cell level correlated with translocated levels of YopB and YopD themselves. Intriguingly, LcrH mutants that are fully competent for effector translocation but produce and translocate lower levels of YopB and YopD also fail to trigger inflammasome activation. Our findings therefore suggest that hypertranslocation of YopD and YopB is linked to inflammasome activation in response to the Yersinia T3SS. IMPORTANCE The innate immune response is critical to effective clearance of pathogens. Recognition of conserved virulence structures and activities by innate immune receptors such as NLRs constitute one of the first steps in mounting the innate immune response. However, pathogens such as Yersinia actively evade or subvert components of host defense, such as inflammasomes. The T3SS-secreted protein YopK is an essential virulence factor that limits translocation of other Yops, thereby limiting T3SS-induced inflammasome activation. However, what triggers inflammasome activation in cells infected by YopK-deficient Yersinia is not clear. Our findings indicate that hypertranslocation of pore complex proteins promotes inflammasome activation and that YopK prevents inflammasome activation by the T3SS by limiting translocation of YopD and YopB themselves. The innate immune response is critical to effective clearance of pathogens. Recognition of conserved virulence structures and activities by innate immune receptors such as NLRs constitute one of the first steps in mounting the innate immune response. However, pathogens such as Yersinia actively evade or subvert components of host defense, such as inflammasomes. The T3SS-secreted protein YopK is an essential virulence factor that limits translocation of other Yops, thereby limiting T3SS-induced inflammasome activation. However, what triggers inflammasome activation in cells infected by YopK-deficient Yersinia is not clear. Our findings indicate that hypertranslocation of pore complex proteins promotes inflammasome activation and that YopK prevents inflammasome activation by the T3SS by limiting translocation of YopD and YopB themselves.

• Publisher: American Society for Microbiology,American Society of Microbiology
• Subject: Animals / Apoptosis / Bacterial Outer Membrane Proteins - metabolism / Blood / Caspase-11 / Cell activation / Cell death / Cell Line / Cell signaling / Cell Survival / Cytokines / Cytotoxicity / Effector cells / Epithelial Cells - microbiology / Epithelial Cells - physiology / Humans / Immune clearance / Immune response / immunologic receptors / Infections / Inflammasomes / Inflammasomes - metabolism / Innate immunity / Kinases / Macrophages / Macrophages - microbiology / Macrophages - physiology / Mice / Microbiology / mikrobiologi / multiprotein complexes / mutants / Mutation / niches / Pathogens / Pattern recognition receptors / Protein Transport / Proteins / Pseudotuberculosis / Salmonella / Sheep / signal transduction / Toxins / type III secretion system / Type III Secretion Systems - metabolism / Virulence / Virulence factors / Yersinia / Yersinia - physiology / Yersinia pseudotuberculosis