scholarly journals Salmonella Pathogenicity Island 2-Encoded Proteins SseC and SseD Are Essential for Virulence and Are Substrates of the Type III Secretion System

2001 ◽  
Vol 69 (2) ◽  
pp. 737-743 ◽  
Author(s):  
Joanna R. Klein ◽  
Bradley D. Jones
Keyword(s):  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Systemic Infection ◽  
Subcellular Location ◽  
Pathogenicity Island ◽  
Secretion System ◽  
Bacterial Membrane ◽  
Phagocytic Cells ◽  
Type Iii ◽  
Bacterial Cell Membrane

ABSTRACT Survival of Salmonella enterica serovar Typhimurium within host phagocytic cells is a critical step in establishing systemic infection in mice. Genes within Salmonellapathogenicity island 2 (SPI-2) encode a type III secretion system that is required for establishment of systemic infection. Several proteins encoded by SPI-2 have homology to type III secreted proteins from enteropathogenic Escherichia coli and Yersiniaand, based on that homology, are predicted to be secreted through the SPI-2 type III secretion system. We have investigated the roles of two of these proteins, SseC and SseD. We demonstrate here that the SseD protein is required for systemic Salmonella infection of the mouse, and we confirmed the virulence requirement for the SseC protein. Experiments were performed, using cellular fractionation and immunoblotting, to identify the subcellular location of the SseC and SseD proteins. Both proteins were found to localize predominantly to the bacterial cell membrane. In addition, our work revealed that SseC and SseD are exposed to the extracellular environment and are loosely associated with the bacterial membrane. Furthermore, localization of SseC and SseD to the bacterial membrane was found to require a functional SPI-2 type III secretion system. Collectively, these results indicate that the SseC and SseD proteins are secreted by the SPI-2 type III secretion system to the bacterial membrane in order to perform their virulence functions.

scholarly journals OmpR Regulates the Two-Component System SsrA-SsrB in Salmonella Pathogenicity Island 2

2000 ◽  
Vol 182 (3) ◽  
pp. 771-781 ◽  
Author(s):  
Anthea K. Lee ◽  
Corrella S. Detweiler ◽  
Stanley Falkow
Keyword(s):  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Regulatory System ◽  
Systemic Infection ◽  
Pathogenicity Island ◽  
Secretion System ◽  
Host Cells ◽  
Type Iii ◽  
Two Component ◽  
Footprint Analysis

ABSTRACT Salmonella pathogenicity island 2 (SPI-2) encodes a putative, two-component regulatory system, SsrA-SsrB, which regulates a type III secretion system needed for replication inside macrophages and systemic infection in mice. The sensor and regulator homologs,ssrAB (spiR), and genes within the secretion system, including the structural gene ssaH, are transcribed after Salmonella enters host cells. We have studied the transcriptional regulation of ssrAB and the secretion system by using gfp fusions to the ssrA andssaH promoters. We found that early transcription ofssrA, after entry into macrophages, is most efficient in the presence of OmpR. An ompR mutant strain does not exhibit replication within cultured macrophages. Furthermore, footprint analysis shows that purified OmpR protein binds directly to thessrA promoter region. We also show that minimal medium, pH 4.5, induces SPI-2 gene expression in wild-type but notompR mutant strains. We conclude that the type III secretion system of SPI-2 is regulated by OmpR, which activates expression of ssrA soon after Salmonella enters the macrophage.

Heterogeneity of type III secretion system (T3SS)-1-independent entry mechanisms used by Salmonella Enteritidis to invade different cell types

Microbiology ◽  
2011 ◽  
Vol 157 (3) ◽  
pp. 839-847 ◽  
Author(s):  
Manon Rosselin ◽  
Nadia Abed ◽  
Isabelle Virlogeux-Payant ◽  
Elisabeth Bottreau ◽  
Pierre-Yves Sizaret ◽  
...  
Keyword(s):  
Cell Lines ◽  
Type Iii Secretion ◽  
Salmonella Enteritidis ◽  
Type Iii Secretion System ◽  
Cell Types ◽  
Secretion System ◽  
Phagocytic Cells ◽  
Type Iii ◽  
Invasion Mechanisms ◽  
Wide Range

Salmonella causes a wide range of diseases from acute gastroenteritis to systemic typhoid fever, depending on the host. To invade non-phagocytic cells, Salmonella has developed different mechanisms. The main invasion system requires a type III secretion system (T3SS) known as T3SS-1, which promotes a Trigger entry mechanism. However, other invasion factors have recently been described in Salmonella, including Rck and PagN, which were not expressed under our bacterial culture conditions. Based on these observations, we used adhesion and invasion assays to analyse the respective roles of Salmonella Enteritidis T3SS-1-dependent and -independent invasion processes at different times of infection. Diverse cell lines and cell types were tested, including endothelial, epithelial and fibroblast cells. We demonstrated that cell susceptibility to the T3SS-1-independent entry differs by a factor of nine between the most and the least permissive cell lines tested. In addition, using scanning electron and confocal microscopy, we showed that T3SS-1-independent entry into cells was characterized by a Trigger-like alteration, as for the T3SS-1-dependent entry, and also by Zipper-like cellular alteration. Our results demonstrate for what is believed to be the first time that Salmonella can induce Trigger-like entry independently of T3SS-1 and can induce Zipper-like entry independently of Rck. Overall, these data open new avenues for discovering new invasion mechanisms in Salmonella.

scholarly journals SseL Is a Salmonella-Specific Translocated Effector Integrated into the SsrB-Controlled Salmonella Pathogenicity Island 2 Type III Secretion System

2006 ◽  
Vol 75 (2) ◽  
pp. 574-580 ◽  
Author(s):  
Brian K. Coombes ◽  
Michael J. Lowden ◽  
Jennifer L. Bishop ◽  
Mark E. Wickham ◽  
Nat F. Brown ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Regulatory System ◽  
Pathogenicity Island ◽  
Secretion System ◽  
Host Cells ◽  
Dependent Manner ◽  
Host Colonization ◽  
Type Iii

ABSTRACT Bacterial pathogens use horizontal gene transfer to acquire virulence factors that influence host colonization, alter virulence traits, and ultimately shape the outcome of disease following infection. One hallmark of the host-pathogen interaction is the prokaryotic type III secretion system that translocates virulence factors into host cells during infection. Salmonella enterica possesses two type III secretion systems that are utilized during host colonization and intracellular replication. Salmonella pathogenicity island 2 (SPI2) is a genomic island containing approximately 30 contiguous genes required to assemble a functional secretion system including the two-component regulatory system called SsrA-SsrB that positively regulates transcription of the secretion apparatus. We used transcriptional profiling with DNA microarrays to search for genes that coregulate with the SPI2 type III secretion machinery in an SsrB-dependent manner. Here we report the identification of a Salmonella-specific translocated effector called SseL that is required for full virulence during murine typhoid-like disease. Analysis of infected macrophages using fluorescence-activated cell sorting revealed that sseL is induced inside cells and requires SsrB for expression. SseL is retained predominantly in the cytoplasm of infected cells following translocation by the type III system encoded in SPI2. Animal infection experiments with sseL mutant bacteria indicate that integration of SseL into the SsrB response regulatory system contributes to systemic virulence of this pathogen.

scholarly journals Fur Regulates Expression of the Salmonella Pathogenicity Island 1 Type III Secretion System through HilD

2007 ◽  
Vol 190 (2) ◽  
pp. 476-486 ◽  
Author(s):  
Jeremy R. Ellermeier ◽  
James M. Slauch
Keyword(s):  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Pathogenicity Island ◽  
Secretion System ◽  
Dependent Manner ◽  
Intestinal Epithelial ◽  
Environmental Signals ◽  
Type Iii ◽  
Regulatory Systems ◽  
Serovar Typhimurium

ABSTRACT The invasion of intestinal epithelial cells by Salmonella enterica serovar Typhimurium is mediated by a type III secretion system (T3SS) encoded on Salmonella pathogenicity island 1 (SPI1). Expression of the SPI1 T3SS is tightly regulated by the combined action of HilC, HilD, and RtsA, three AraC family members that can independently activate hilA, which encodes the direct regulator of the SPI1 structural genes. Expression of hilC, hilD, and rtsA is controlled by a number of regulators that respond to a variety of environmental signals. In this work, we show that one such signal is iron mediated by Fur (ferric uptake regulator). Fur activates hilA transcription in a HilD-dependent manner. Fur regulation of HilD does not appear to be simply at the transcriptional or translational level but rather requires the presence of the HilD protein. Fur activation of SPI1 is not mediated through the Fur-regulated small RNAs RfrA and RfrB, which are the Salmonella ortholog and paralog of RyhB that control expression of sodB. Fur regulation of HilD is also not mediated through the known SPI1 repressor HilE or the CsrABC system. Although understanding the direct mechanism of Fur action on HilD requires further analysis, this work is an important step toward elucidating how various global regulatory systems control SPI1.

scholarly journals Differential expression of Salmonella type III secretion system factors InvJ, PrgJ, SipC, SipD, SopA and SopB in cultures and in mice

Microbiology ◽  
2010 ◽  
Vol 156 (1) ◽  
pp. 116-127 ◽  
Author(s):  
Hao Gong ◽  
Gia-Phong Vu ◽  
Yong Bai ◽  
Edward Yang ◽  
Fenyong Liu ◽  
...  
Keyword(s):  
Differential Expression ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Systemic Infection ◽  
Secretion System ◽  
Flag Epitope ◽  
Type Iii ◽  
Late Stages

The type III secretion system (T3SS) encoded by Salmonella pathogenicity island 1 (SPI-1) is important for the invasion of epithelial cells during development of Salmonella-associated enterocolitis. It has been suggested that the level and timing of the expression of the SPI-1 T3SS proteins and effectors dictate the consequences of bacterial infection and pathogenesis. However, the expression of these proteins has not been extensively studied in vivo, especially during the later stages of salmonellosis when the infection is established. We have constructed recombinant Salmonella strains that contain a FLAG epitope inserted in-frame to genes invJ, prgJ, sipC, sipD, sopA and sopB, and investigated the expression of the tagged proteins both in vitro and in vivo during murine salmonellosis. Mice were inoculated intraperitoneally or intragastrically with the tagged Salmonella strains. At different time points post-infection, bacteria were recovered from various organs, and the expression of the tagged proteins was determined. Our results provide direct evidence that PrgJ and SipD are expressed in Salmonella colonizing the liver and ileum of infected animals at both the early and late stages of infection. Furthermore, our study has shown that the InvJ protein is expressed preferentially in Salmonella colonizing the ileum but not the liver, while SipC is expressed preferentially in Salmonella colonizing the liver but not the ileum. Thus, Salmonella appears to express different SPI-1 proteins and effectors when colonizing specific tissues. Our results suggest that differential expression of these proteins may be important for tissue-specific aspects of bacterial pathogenesis such as gastroenterititis in the ileum and systemic infection in the liver.

Attenuated virulence of an Aeromonas salmonicida subsp. salmonicida type III secretion mutant in a rainbow trout model

Microbiology ◽  
2005 ◽  
Vol 151 (6) ◽  
pp. 2111-2118 ◽  
Author(s):  
Sarah E. Burr ◽  
Dmitri Pugovkin ◽  
Thomas Wahli ◽  
Helmut Segner ◽  
Joachim Frey
Keyword(s):  
Rainbow Trout ◽  
Histological Examination ◽  
Type Iii Secretion ◽  
Systemic Disease ◽  
Type Iii Secretion System ◽  
Systemic Infection ◽  
Secretion System ◽  
Aeromonas Salmonicida ◽  
Blood Leukocytes ◽  
Type Iii

Aeromonas salmonicida subsp. salmonicida is the causative agent of furunculosis, a severe systemic disease affecting salmonid fish. This bacterium contains a type III protein secretion system that is responsible for the secretion and translocation of the ADP-ribosylating toxin, AexT, into the cytosol of fish cells. This study showed that inactivation of the type III secretion system by marker-replacement mutagenesis of the gene ascV, which encodes an inner-membrane component of the type III secretion system, attenuated virulence in a rainbow trout model. The isogenic ascV deletion mutant was phagocytosed by peripheral blood leukocytes but the wild-type (wt) A. salmonicida subsp. salmonicida isolate was not. Histological examination of fish experimentally infected with the wt bacterium revealed extensive tissue necrosis and bacterial aggregates in all organs examined, including the heart, kidney and liver, indicating that the isolate established a systemic infection. Cumulative mortality of fish experimentally infected with the wt bacterium reached 88 %. In contrast, no mortality was observed among fish infected with the same dose of the ascV mutant, and histological examination of fish infected with this strain revealed healthy organs. The results indicate that the type III secretion system of A. salmonicida subsp. salmonicida is required to establish systemic infection.

scholarly journals Regulation of the Edwardsiella ictaluri Type III Secretion System by pH and Phosphate Concentration through EsrA, EsrB, and EsrC

2011 ◽  
Vol 77 (13) ◽  
pp. 4293-4302 ◽  
Author(s):  
Matthew L. Rogge ◽  
Ronald L. Thune
Keyword(s):  
Channel Catfish ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Pathogenicity Island ◽  
Low Ph ◽  
Secretion System ◽  
Edwardsiella Ictaluri ◽  
Phosphate Limitation ◽  
Content Type ◽  
Type Iii

ABSTRACTA recently describedEdwardsiella ictaluritype III secretion system (T3SS) with functional similarity to theSalmonellapathogenicity island 2 T3SS is required for replication in channel catfish head-kidney-derived macrophages (HKDM) and virulence in channel catfish. Quantitative PCR and Western blotting identified low pH and phosphate limitation as conducive to expression of theE. ictaluriT3SS, growth conditions that mimic the phagosomal environment. Mutagenesis studies demonstrated that expression is under the control of the EsrAB two-component regulatory system. EsrB also induces upregulation of the AraC-type regulatory protein EsrC, which enhances expression of the EscB/EseG chaperone/effector operon in concert with EsrB and induces expression of the pEI1-encoded effector, EseH. EsrC also induces expression of a putative type VI secretion system translocon protein, EvpC, which is secreted under the same low-pH conditions as the T3SS translocon proteins. The pEI2-encoded effector, EseI, was upregulated under low-pH and low-phosphate conditions but not in an EsrB- or EsrC-dependent manner. Mutations of EsrA and EsrB both resulted in loss of the ability to replicate in HKDM and full attenuation in the channel catfish host. Mutation of EsrC did not affect intracellular replication but did result in attenuation in catfish. Although EsrB is the primary transcriptional regulator forE. ictalurigenes within the T3SS pathogenicity island, EsrC regulates expression of the plasmid-carried effectoreseHand appears to mediate coordinated expression of the T6SS with the T3SS.

SseA is a chaperone for the SseB and SseD translocon components of the Salmonella pathogenicity-island-2-encoded type III secretion system

Microbiology ◽  
2003 ◽  
Vol 149 (5) ◽  
pp. 1103-1111 ◽  
Author(s):  
Javier Ruiz-Albert ◽  
Rosanna Mundy ◽  
Xiu-Jun Yu ◽  
Carmen R. Beuzón ◽  
David W. Holden
Keyword(s):  
Type Iii Secretion ◽  
Sequence Similarity ◽  
Coiled Coil ◽  
Type Iii Secretion System ◽  
Pathogenicity Island ◽  
Secretion System ◽  
Effector Proteins ◽  
Type Iii ◽  
Virulence Attenuation

The type III secretion system (TTSS) encoded by the Salmonella pathogenicity island 2 (SPI-2) is required for bacterial replication inside macrophages and for systemic infection in mice. Many TTSS secreted proteins, including effectors and components of the translocon, require chaperones which promote their stability, prevent their premature interactions or facilitate their secretion. In this study, the function of the first gene (sseA) of one of the SPI-2 operons (sseA–G) was investigated. This operon includes genes that encode translocon components (SseB, SseC and SseD), translocated proteins (SseF and SseG) and putative chaperones (SscA and SscB). sseA encodes a 12·5 kDa protein with a C-terminal region with the potential to form a coiled-coil structure, but no sequence similarity to other proteins. Mutation of sseA results in severe virulence attenuation and an intracellular replication defect. It is shown here that SseA is not a secreted protein, but is required for SPI-2-dependent translocation of two effector proteins (SifA and PipB). Furthermore, the translocon components SseB and SseD were not detected in an sseA mutant strain. By using a yeast two-hybrid assay and column binding experiments, it is demonstrated that SseA interacts directly with SseB and SseD. These results indicate that SseA is a chaperone for SseB and SseD. The inability of an sseA mutant to assemble the SPI-2 TTSS translocon accounts for its high level of virulence attenuation in vivo. To the authors' knowledge, this is the first chaperone described for the SPI-2 TTSS.

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