C-terminal amino acid residues of the trimeric autotransporter adhesin YadA of Yersinia enterocolitica are decisive for its recognition and assembly by BamA

Lehr, U.; Schütz, Monika; Oberhettinger, Philipp; Ruı́z-Pérez, Fernando; Donald, J; Palmer, Tracy; Linke, Dirk; Henderson, Ian R.; Autenrieth, Ingo B.

doi:10.1111/j.1365-2958.2010.07377.x

Cited by 79 publications

(102 citation statements)

References 46 publications

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“…Unassembled ␤-barrel OMPs are frequently eliminated from the periplasm by the protease activity of DegP (3,26,28). To assess whether unfolded FimD in SurA-depleted cells is degraded by DegP, FimD levels were analyzed in a double conditional mutant strain, UTdegP-P BAD ::surA (Table 1), which carries a deletion of degP and the surA gene under the control of the P BAD promoter.…”

Section: Resultsmentioning

confidence: 99%

The Fimbrial Usher FimD Follows the SurA-BamB Pathway for Its Assembly in the Outer Membrane of Escherichia coli

2011

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Fimbrial ushers are the largest ␤-barrel outer membrane proteins (OMPs) known to date, which function in the polymerization of fimbriae and their translocation to the bacterial surface. Folding and assembly of these complex OMPs are not characterized. Here, we investigate the role of periplasmic chaperones (SurA, Skp, DegP, and FkpA) and individual components of the ␤-barrel assembly machinery (BAM) complex (BamA, BamB, BamC, and BamE) in the folding of the Escherichia coli FimD usher. The FimD level is dramatically reduced (ϳ30-fold) in a surA null mutant, but a strong cell envelope stress is constitutively activated with upregulation of DegP (ϳ10-fold). To demonstrate a direct role of SurA, FimD folding was analyzed in a conditional surA mutant in which SurA expression was controlled. In this strain, FimD is depleted from bacteria in parallel to SurA without significant upregulation of DegP. Interestingly, the dependency on SurA is higher for FimD than for other OMPs. We also demonstrate that a functional BAM complex is needed for folding of FimD. In addition, FimD levels were strongly reduced (ϳ5-fold) in a mutant lacking the accessory lipoprotein BamB. The critical role of BamB for FimD folding was confirmed by complementation and BamB depletion experiments. Similar to SurA dependency, FimD showed a stronger dependency on BamB than OMPs. On the other hand, folding of FimD was only marginally affected in bamC and bamE mutants. Collectively, our results indicate that FimD usher follows the SurA-BamB pathway for its assembly. The preferential use of this pathway for the folding of OMPs with large ␤-barrels is discussed.Gram-negative bacteria display on their surfaces adhesive multimeric protein fibers known as pili, fimbriae, or fibrillae that mediate attachment to host cells and tissues for colonization and infection (12,24). The assembly of a large family of these appendages depends on the conserved chaperone-usher (CU) pathway (36, 49). The pilin subunits that form these fibers initially cross the cytoplasmic membrane through the Sec translocon to reach the periplasmic space, where they bind to their cognate chaperones, forming binary complexes. These complexes are specifically targeted to the usher, a ϳ90-kDa integral outer membrane protein (OMP), that catalyzes the polymerization of pilin subunits in an ordered manner and secretes the resulting protein fiber to the bacterial surface (32,34,37). Type 1 (fim) and P (pap) fimbriae from uropathogenic Escherichia coli (UPEC) strains have been established as paradigms of the CU pathway, and their respective chaperones and ushers are studied as models of these types of proteins. Biochemical and structural studies of the usher proteins of type 1 and P fimbriae (named FimD and PapC, respectively) indicate that these proteins may form dimers in the OM and contain the largest ␤-barrel known to date in OMPs, with 24 antiparallel ␤-strands (18,34,37). In addition to the large ␤-barrel domain, FimD and PapC ushers contain several globular domains, including periplasmic ...

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Section: Resultsmentioning

confidence: 99%

The Fimbrial Usher FimD Follows the SurA-BamB Pathway for Its Assembly in the Outer Membrane of Escherichia coli

2011

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“…It is worth noting that the trimeric autotransporters also depend on BamA for membrane insertion and also seem to form hairpin intermediates (34,44,49). It is hard to conceive how three hairpins could be formed and how three parts of a transmembrane ␤-barrel could be assembled and inserted simultaneously by one BAM complex, and it is difficult to imagine three individual BAM complexes working in a synchronous fashion.…”

Section: Discussionmentioning

confidence: 99%

The Inverse Autotransporter Intimin Exports Its Passenger Domain via a Hairpin Intermediate

Oberhettinger

Leo

Linke

et al. 2015

Journal of Biological Chemistry

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Background:Intimin exports its C-terminal passenger domain through an N-terminal ␤-barrel onto the bacterial surface. Results: Insertion of an epitope tag at the very N terminus of the passenger domain stalls the export of the passenger. Conclusion:The intimin passenger adopts a hairpin conformation during translocation. Significance: Our results confirm the hairpin model of inverse autotransport where the passenger is translocated from the N to the C terminus.

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“…However, the assistance of the Bam complex (previously known as Omp85 or YaeT) is required in both (16,20,22,25,40). Strains with deletion of Omp85 were not able to transport IgA protease, a classical autotransporter.…”

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confidence: 99%

The Translocation Domain in Trimeric Autotransporter Adhesins Is Necessary and Sufficient for Trimerization and Autotransportation

et al. 2012

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Trimeric autotransporter adhesins (TAAs) comprise one of the secretion pathways of the type V secretion system. The mechanism of their translocation across the outer membrane remains unclear, but it most probably occurs by the formation of a hairpin inside the ␤-barrel translocation unit, leading to transportation of the passenger domain from the C terminus to the N terminus through the lumen of the ␤-barrel. We further investigated the phenomenon of autotransportation and the rules that govern it. We showed by coexpressing different Escherichia coli immunoglobulin-binding (Eib) proteins that highly similar TAAs could form stochastically mixed structures (heterotrimers). We further investigated this phenomenon by coexpressing two more distantly related TAAs, EibA and YadA. These, however, did not form heterotrimers; indeed, coexpression was lethal to the cells, leading to elimination of one or another of the genes. However, substituting in either protein the barrel of the other one so that the barrels were identical led to formation of heterotrimers as for Eibs. Our work shows that trimerization of the ␤-barrel, but not the passenger domain, is necessary and sufficient for TAA secretion while the passenger domain is not.

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C-terminal amino acid residues of the trimeric autotransporter adhesin YadA of Yersinia enterocolitica are decisive for its recognition and assembly by BamA

Cited by 79 publications

References 46 publications

The Fimbrial Usher FimD Follows the SurA-BamB Pathway for Its Assembly in the Outer Membrane of Escherichia coli

The Fimbrial Usher FimD Follows the SurA-BamB Pathway for Its Assembly in the Outer Membrane of Escherichia coli

The Inverse Autotransporter Intimin Exports Its Passenger Domain via a Hairpin Intermediate

The Translocation Domain in Trimeric Autotransporter Adhesins Is Necessary and Sufficient for Trimerization and Autotransportation

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