Deletion of the genes encoding the type III secretion system and cytotoxic enterotoxin alters host responses to Aeromonas hydrophila infection

2006 ◽  
Vol 40 (5) ◽  
pp. 198-210 ◽  
Author(s):  
Amin A. Fadl ◽  
Cristi L. Galindo ◽  
Jian Sha ◽  
Tatiana E. Erova ◽  
Clifford W. Houston ◽  
...  
Keyword(s):  
Aeromonas Hydrophila ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Host Responses ◽  
Type Iii ◽  
Genes Encoding

scholarly journals Expression of the Bradyrhizobium japonicum Type III Secretion System in Legume Nodules and Analysis of the Associated tts box Promoter

2008 ◽  
Vol 21 (8) ◽  
pp. 1087-1093 ◽  
Author(s):  
Susanne Zehner ◽  
Grit Schober ◽  
Mandy Wenzel ◽  
Kathrin Lang ◽  
Michael Göttfert
Keyword(s):  
Type Iii Secretion ◽  
Bradyrhizobium Japonicum ◽  
Response Regulator ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Symbiotic Efficiency ◽  
Type Iii ◽  
Genes Encoding ◽  
Transcriptional Start Sites ◽  
Type Three Secretion

In Bradyrhizobium japonicum, as in some other rhizobia, symbiotic efficiency is influenced by a type III secretion system (T3SS). Most genes encoding the transport machinery and secreted proteins are preceded by a conserved 30-bp motif, the type-three secretion (tts) box. In this study, we found that regions downstream of 34 tts boxes are transcribed. For nopB, nopL, and gunA2, the transcriptional start sites were found to be 12, 11, and 10 bp downstream of their tts boxes, respectively. The deletion of this motif or modification of two or more conserved residues strongly reduced expression of nopB. This indicates that the tts box is an essential promoter element. Data obtained with lacZ reporter gene fusions of five genes preceded by a tts box (gunA2, nopB, rhcV, nopL, and blr1806) revealed that they are expressed in 4-week-old nodules of Macroptilium atropurpureum. These data suggest that the T3SS is active in mature nitrogen-fixing nodules. The two-component response regulator TtsI is required for the expression of rhcV, nopL, and blr1806 in bacteroids. Staining of inoculated roots showed that nopB is also expressed in early infection stages.

scholarly journals The Type III Secretion System and Cytotoxic Enterotoxin Alter the Virulence of Aeromonas hydrophila

2005 ◽  
Vol 73 (10) ◽  
pp. 6446-6457 ◽  
Author(s):  
Jian Sha ◽  
Lakshmi Pillai ◽  
Amin A. Fadl ◽  
Cristi L. Galindo ◽  
Tatiana E. Erova ◽  
...  
Keyword(s):  
Aeromonas Hydrophila ◽  
Type Iii Secretion ◽  
Cultured Cells ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Effector Proteins ◽  
Host Cells ◽  
Gene Encoding ◽  
Type Iii ◽  
Isogenic Mutants

ABSTRACT Many gram-negative bacteria use a type III secretion system (TTSS) to deliver effector proteins into host cells. Here we report the characterization of a TTSS chromosomal operon from the diarrheal isolate SSU of Aeromonas hydrophila. We deleted the gene encoding Aeromonas outer membrane protein B (AopB), which is predicted to be involved in the formation of the TTSS translocon, from wild-type (WT) A. hydrophila as well as from a previously characterized cytotoxic enterotoxin gene (act)-minus strain of A. hydrophila, thus generating aopB and act/aopB isogenic mutants. The act gene encodes a type II-secreted cytotoxic enterotoxin (Act) that has hemolytic, cytotoxic, and enterotoxic activities and induces lethality in a mouse model. These isogenic mutants (aopB, act, and act/aopB) were highly attenuated in their ability to induce cytotoxicity in RAW 264.7 murine macrophages and HT-29 human colonic epithelial cells. The act/aopB mutant demonstrated the greatest reduction in cytotoxicity to cultured cells after 4 h of infection, as measured by the release of lactate dehydrogenase enzyme, and was avirulent in mice, with a 90% survival rate compared to that of animals infected with Act and AopB mutants, which caused 50 to 60% of the animals to die at a dose of three 50% lethal doses. In contrast, WT A. hydrophila killed 100% of the mice within 48 h. The effects of these mutations on cytotoxicity could be complemented with the native genes. Our studies further revealed that the production of lactones, which are involved in quorum sensing (QS), was decreased in the act (32%) and aopB (64%) mutants and was minimal (only 8%) in the act/aopB mutant, compared to that of WT A. hydrophila SSU. The effects of act and aopB gene deletions on lactone production could also be complemented with the native genes, indicating specific effects of Act and the TTSS on lactone production. Although recent studies with other bacteria have indicated TTSS regulation by QS, this is the first report describing a correlation between the TTSS and Act of A. hydrophila and the production of lactones.

Evaluation of biofilm production and characterization of genes encoding type III secretion system among Pseudomonas aeruginosa isolated from burn patients

Burns ◽  
2012 ◽  
Vol 38 (8) ◽  
pp. 1192-1197 ◽  
Author(s):  
Fereshteh Jabalameli ◽  
Akbar Mirsalehian ◽  
Babak Khoramian ◽  
Marzieh Aligholi ◽  
Seyed Sajjad Khoramrooz ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Burn Patients ◽  
Type Iii ◽  
Biofilm Production ◽  
Genes Encoding

scholarly journals A Type III Secretion System Is Required for Aeromonas hydrophila AH-1 Pathogenesis

2004 ◽  
Vol 72 (3) ◽  
pp. 1248-1256 ◽  
Author(s):  
H. B. Yu ◽  
P. S. Srinivasa Rao ◽  
H. C. Lee ◽  
S. Vilches ◽  
S. Merino ◽  
...  
Keyword(s):  
Aeromonas Hydrophila ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Gene Clusters ◽  
Opportunistic Pathogen ◽  
Secretion System ◽  
Virulence Determinants ◽  
Degenerate Primers ◽  
Type Iii ◽  
Insertional Inactivation

ABSTRACT Aeromonas hydrophila is a gram-negative opportunistic pathogen in fish and humans. Many bacterial pathogens of animals and plants have been shown to inject anti-host virulence determinants into the hosts via a type III secretion system (TTSS). Degenerate primers based on lcrD family genes that are present in every known TTSS allowed us to locate the TTSS gene cluster in A. hydrophila AH-1. A series of genome walking steps helped in the identification of 25 open reading frames that encode proteins homologous to those in TTSSs in other bacteria. PCR-based analysis showed the presence of lcrD homologs (ascV) in all of the 33 strains of A. hydrophila isolated from various sources. Insertional inactivation of two of the TTSS genes (aopB and aopD) led to decreased cytotoxicity in carp epithelial cells, increased phagocytosis, and reduced virulence in blue gourami. These results show that a TTSS is required for A. hydrophila pathogenesis. This is the first report of sequencing and characterization of TTSS gene clusters from A. hydrophila. The TTSS identified here may help in developing suitable vaccines as well as in further understanding of the pathogenesis of A. hydrophila.

scholarly journals Chlamydial Type III Secretion System Is Encoded on Ten Operons Preceded by Sigma 70-Like Promoter Elements

2006 ◽  
Vol 189 (1) ◽  
pp. 198-206 ◽  
Author(s):  
P. Scott Hefty ◽  
Richard S. Stephens
Keyword(s):  
Type Iii Secretion ◽  
Transcriptional Start Site ◽  
Type Iii Secretion System ◽  
Regulatory Elements ◽  
Secretion System ◽  
Secretion Systems ◽  
Promoter Elements ◽  
Type Iii ◽  
Type Iii Secretion Systems ◽  
Genes Encoding

ABSTRACT Many gram-negative bacterial pathogens employ type III secretion systems for infectious processes. Chlamydiae are obligate intracellular bacteria that encode a conserved type III secretion system that is likely requisite for growth. Typically, genes encoding type III secretion systems are located in a single locus; however, for chlamydiae these genes are scattered throughout the genome. Little is known regarding the gene regulatory mechanisms for this essential virulence determinant. To facilitate identification of cis-acting transcriptional regulatory elements, the operon structure was determined. This analysis revealed 10 operons that contained 37 genes associated with the type III secretion system. Linkage within these operons suggests a role in type III secretion for each of these genes, including 13 genes encoding proteins with unknown function. The transcriptional start site for each operon was determined. In conjunction with promoter activity assays, this analysis revealed that the type III secretion system operons encode σ70-like promoter elements. Transcriptional initiation by a sigma factor responsible for constitutive gene expression indicates that undefined activators or repressors regulate developmental stage-specific expression of chlamydial type III secretion system genes.

scholarly journals Mutations in Salmonella Pathogenicity Island 2 (SPI2) Genes Affecting Transcription of SPI1 Genes and Resistance to Antimicrobial Agents

1998 ◽  
Vol 180 (18) ◽  
pp. 4775-4780 ◽  
Author(s):  
Jörg Deiwick ◽  
Thomas Nikolaus ◽  
Jaqueline E. Shea ◽  
Colin Gleeson ◽  
David W. Holden ◽  
...  
Keyword(s):  
Type Iii Secretion ◽  
Antimicrobial Agents ◽  
Type Iii Secretion System ◽  
Polymyxin B ◽  
Secretion System ◽  
Effector Proteins ◽  
Secretion Systems ◽  
Type Iii ◽  
Genes Encoding

ABSTRACT The Salmonella typhimurium genome contains two pathogenicity islands (SPI) with genes encoding type III secretion systems for virulence proteins. SPI1 is required for the penetration of the epithelial layer of the intestine. SPI2 is important for the subsequent proliferation of bacteria in the spleens of infected hosts. Although most mutations in SPI2 lead to a strong reduction of virulence, they have different effects in vitro, with some mutants having significantly increased sensitivity to gentamicin and the antibacterial peptide polymyxin B. Previously we showed that certain mutations in SPI2 affect the ability of S. typhimurium to secrete SPI1 effector proteins and to invade cultured eukaryotic cells. In this study, we show that these SPI2 mutations affect the expression of the SPI1 invasion genes. Analysis of reporter fusions to various SPI1 genes reveals highly reduced expression of sipC,prgK, and hilA, the transcriptional activator of SPI1 genes. These observations indicate that the expression of one type III secretion system can be influenced dramatically by mutations in genes encoding a second type III secretion system in the same cell.

scholarly journals Complete Genome Sequence of Uropathogenic Proteus mirabilis, a Master of both Adherence and Motility

2008 ◽  
Vol 190 (11) ◽  
pp. 4027-4037 ◽  
Author(s):  
Melanie M. Pearson ◽  
Mohammed Sebaihia ◽  
Carol Churcher ◽  
Michael A. Quail ◽  
Aswin S. Seshasayee ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Proteus Mirabilis ◽  
Complete Genome Sequence ◽  
Type Iii Secretion ◽  
Complete Genome ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Type Iii ◽  
Genes Encoding

ABSTRACT The gram-negative enteric bacterium Proteus mirabilis is a frequent cause of urinary tract infections in individuals with long-term indwelling catheters or with complicated urinary tracts (e.g., due to spinal cord injury or anatomic abnormality). P. mirabilis bacteriuria may lead to acute pyelonephritis, fever, and bacteremia. Most notoriously, this pathogen uses urease to catalyze the formation of kidney and bladder stones or to encrust or obstruct indwelling urinary catheters. Here we report the complete genome sequence of P. mirabilis HI4320, a representative strain cultured in our laboratory from the urine of a nursing home patient with a long-term (≥30 days) indwelling urinary catheter. The genome is 4.063 Mb long and has a G+C content of 38.88%. There is a single plasmid consisting of 36,289 nucleotides. Annotation of the genome identified 3,685 coding sequences and seven rRNA loci. Analysis of the sequence confirmed the presence of previously identified virulence determinants, as well as a contiguous 54-kb flagellar regulon and 17 types of fimbriae. Genes encoding a potential type III secretion system were identified on a low-G+C-content genomic island containing 24 intact genes that appear to encode all components necessary to assemble a type III secretion system needle complex. In addition, the P. mirabilis HI4320 genome possesses four tandem copies of the zapE metalloprotease gene, genes encoding six putative autotransporters, an extension of the atf fimbrial operon to six genes, including an mrpJ homolog, and genes encoding at least five iron uptake mechanisms, two potential type IV secretion systems, and 16 two-component regulators.

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