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Updated: Jun 10, 2026

Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
Published on: December 17, 2013
The type 2 secretion Pseudopilin, gspJ, is required for multihost pathogenicity of Burkholderia cenocepacia AU1054
Vishal S Somvanshi1, Poorna Viswanathan, Janette L Jacobs
1Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USA.
Abstract:
Burkholderia cenocepacia AU1054 is an opportunistic pathogen isolated from the blood of a person with cystic fibrosis. AU1054 is a multihost pathogen causing rapid pathogenicity to Caenorhabditis elegans nematodes. Within 24 h, AU1054 causes greater than 50% mortality, reduced growth, emaciated body, distended intestinal lumen, rectal swelling, and prolific infection of the nematode intestine. To determine virulence mechanisms, 3,000 transposon mutants were screened for attenuated virulence in nematodes. Fourteen virulence-attenuated mutants were isolated, and the mutant genes were identified. These genes included paaA, previously identified as being required for full virulence of B. cenocepacia K56-2. Six mutants were restored in virulence by complementation with their respective wild-type gene. One of these contained an insertion in gspJ, predicted to encode a pseudopilin component of the type 2 secretion system (T2SS). Nematodes infected with AU1054 gspJ had fewer bacteria present in the intestine than those infected with the wild type but still showed rectal swelling. The gspJ mutant was also defective in pathogenicity to onion and in degradation of polygalacturonic acid and casein. This result differs from previous studies where no or little role was found for T2SS in Burkholderia virulence, although virulence factors such as zinc metalloproteases and polygalacturonase are known to be secreted by the T2SS. This study highlights strain specific differences in B. cenocepacia virulence mechanisms important for understanding what enables environmental microbes to function as opportunistic pathogens.
Insights
Burkholderia cenocepacia AU1054 uses a type 2 secretion system (T2SS) for virulence, unlike other strains. This study identifies key virulence factors in this opportunistic pathogen, crucial for understanding cystic fibrosis infections.
Area of Science:
- Microbiology
- Pathogenesis
- Bacterial Genetics
Background:
- Burkholderia cenocepacia AU1054 is an opportunistic pathogen linked to cystic fibrosis.
- This strain exhibits rapid pathogenicity in Caenorhabditis elegans nematodes, causing mortality and distinct physiological damage within 24 hours.
Purpose of the Study:
- To identify the virulence mechanisms of Burkholderia cenocepacia AU1054.
- To investigate the role of the type 2 secretion system (T2SS) in the pathogenicity of this specific strain.
Main Methods:
- Screening of 3,000 transposon mutants of B. cenocepacia AU1054 for attenuated virulence in C. elegans.
- Identification of mutated genes in virulence-attenuated mutants.
- Complementation assays to confirm gene function.
- Assessment of pathogenicity in other hosts (onion) and substrate degradation (polygalacturonic acid, casein).
Main Results:
- Fourteen virulence-attenuated mutants were identified, including mutations in paaA and gspJ.
- A gspJ mutant, encoding a T2SS component, showed reduced bacterial load in nematodes but retained rectal swelling.
- The gspJ mutant exhibited defects in onion pathogenicity and degradation of polygalacturonic acid and casein.
Conclusions:
- The type 2 secretion system (T2SS) plays a significant role in the virulence of Burkholderia cenocepacia AU1054, contrary to previous findings in other strains.
- Strain-specific differences in virulence mechanisms are critical for understanding how environmental bacteria become opportunistic pathogens.
- Identification of virulence factors like those secreted by T2SS is essential for therapeutic strategies against B. cenocepacia infections.
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