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

U. Lehr, M. Schuetz, P. Oberhettinger, F. Ruiz-Perez, J. W. Donald, T. Palmer, D. Linke, I. R. Henderson, I. B. Autenrieth

    Research output: Contribution to journalArticle

    56 Citations (Scopus)

    Abstract

    P>The Bam complex is a highly conserved multiprotein machine essential for the assembly of beta-barrel outer membrane proteins. It is composed of the essential outer membrane protein BamA and four outer membrane associated lipoproteins BamB-E. The Yersinia enterocolitica Adhesin A (YadA) is the prototype of trimeric auotransporter adhesins (TAAs), consisting of a head, stalk and a beta-barrel membrane anchor. To investigate the role of BamA in biogenesis of TAAs, we expressed YadA in a BamA-depleted strain of Escherichia coli, which resulted in degradation of YadA. Yeast-two-hybrid experiments and immunofluorescence studies revealed that BamA and YadA interact directly and colocalize. As BamA recognizes the C-terminus of OMPs, we exchanged the nine most C-terminal amino acids of YadA. Substitution of the amino acids in position 1, 3 or 5 from the C-terminus with glycine resulted in DegP-dependent degradation of YadA. Despite degradation all YadA proteins assembled in the outer membrane. In summary we demonstrate that (i) BamA is essential for biogenesis of the TAA YadA, (ii) BamA interacts directly with YadA, (iii) the C-terminal amino acid motif of YadA is important for the BamA-dependent assembly and differs slightly compared with other OMPs, and (iv) BamA and YadA colocalize.

    Original languageEnglish
    Pages (from-to)932-946
    Number of pages15
    JournalMolecular Microbiology
    Volume78
    Issue number4
    DOIs
    Publication statusPublished - 2010

    Keywords

    • OUTER-MEMBRANE-PROTEIN
    • CONSERVED BACTERIAL PROTEIN
    • ESCHERICHIA-COLI
    • VIRULENCE PLASMID
    • BINDING DOMAIN
    • OMP85
    • SECRETION
    • PATHWAY
    • TRANSLOCATION
    • TRANSPORT

    Cite this

    Lehr, U. ; Schuetz, M. ; Oberhettinger, P. ; Ruiz-Perez, F. ; Donald, J. W. ; Palmer, T. ; Linke, D. ; Henderson, I. R. ; Autenrieth, I. B. / C-terminal amino acid residues of the trimeric autotransporter adhesin YadA of Yersinia enterocolitica are decisive for its recognition and assembly by BamA. In: Molecular Microbiology. 2010 ; Vol. 78, No. 4. pp. 932-946.
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    title = "C-terminal amino acid residues of the trimeric autotransporter adhesin YadA of Yersinia enterocolitica are decisive for its recognition and assembly by BamA",
    abstract = "P>The Bam complex is a highly conserved multiprotein machine essential for the assembly of beta-barrel outer membrane proteins. It is composed of the essential outer membrane protein BamA and four outer membrane associated lipoproteins BamB-E. The Yersinia enterocolitica Adhesin A (YadA) is the prototype of trimeric auotransporter adhesins (TAAs), consisting of a head, stalk and a beta-barrel membrane anchor. To investigate the role of BamA in biogenesis of TAAs, we expressed YadA in a BamA-depleted strain of Escherichia coli, which resulted in degradation of YadA. Yeast-two-hybrid experiments and immunofluorescence studies revealed that BamA and YadA interact directly and colocalize. As BamA recognizes the C-terminus of OMPs, we exchanged the nine most C-terminal amino acids of YadA. Substitution of the amino acids in position 1, 3 or 5 from the C-terminus with glycine resulted in DegP-dependent degradation of YadA. Despite degradation all YadA proteins assembled in the outer membrane. In summary we demonstrate that (i) BamA is essential for biogenesis of the TAA YadA, (ii) BamA interacts directly with YadA, (iii) the C-terminal amino acid motif of YadA is important for the BamA-dependent assembly and differs slightly compared with other OMPs, and (iv) BamA and YadA colocalize.",
    keywords = "OUTER-MEMBRANE-PROTEIN, CONSERVED BACTERIAL PROTEIN, ESCHERICHIA-COLI, VIRULENCE PLASMID, BINDING DOMAIN, OMP85, SECRETION, PATHWAY, TRANSLOCATION, TRANSPORT",
    author = "U. Lehr and M. Schuetz and P. Oberhettinger and F. Ruiz-Perez and Donald, {J. W.} and T. Palmer and D. Linke and Henderson, {I. R.} and Autenrieth, {I. B.}",
    year = "2010",
    doi = "10.1111/j.1365-2958.2010.07377.x",
    language = "English",
    volume = "78",
    pages = "932--946",
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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. / Lehr, U.; Schuetz, M.; Oberhettinger, P.; Ruiz-Perez, F.; Donald, J. W.; Palmer, T.; Linke, D.; Henderson, I. R.; Autenrieth, I. B.

    In: Molecular Microbiology, Vol. 78, No. 4, 2010, p. 932-946.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - C-terminal amino acid residues of the trimeric autotransporter adhesin YadA of Yersinia enterocolitica are decisive for its recognition and assembly by BamA

    AU - Lehr, U.

    AU - Schuetz, M.

    AU - Oberhettinger, P.

    AU - Ruiz-Perez, F.

    AU - Donald, J. W.

    AU - Palmer, T.

    AU - Linke, D.

    AU - Henderson, I. R.

    AU - Autenrieth, I. B.

    PY - 2010

    Y1 - 2010

    N2 - P>The Bam complex is a highly conserved multiprotein machine essential for the assembly of beta-barrel outer membrane proteins. It is composed of the essential outer membrane protein BamA and four outer membrane associated lipoproteins BamB-E. The Yersinia enterocolitica Adhesin A (YadA) is the prototype of trimeric auotransporter adhesins (TAAs), consisting of a head, stalk and a beta-barrel membrane anchor. To investigate the role of BamA in biogenesis of TAAs, we expressed YadA in a BamA-depleted strain of Escherichia coli, which resulted in degradation of YadA. Yeast-two-hybrid experiments and immunofluorescence studies revealed that BamA and YadA interact directly and colocalize. As BamA recognizes the C-terminus of OMPs, we exchanged the nine most C-terminal amino acids of YadA. Substitution of the amino acids in position 1, 3 or 5 from the C-terminus with glycine resulted in DegP-dependent degradation of YadA. Despite degradation all YadA proteins assembled in the outer membrane. In summary we demonstrate that (i) BamA is essential for biogenesis of the TAA YadA, (ii) BamA interacts directly with YadA, (iii) the C-terminal amino acid motif of YadA is important for the BamA-dependent assembly and differs slightly compared with other OMPs, and (iv) BamA and YadA colocalize.

    AB - P>The Bam complex is a highly conserved multiprotein machine essential for the assembly of beta-barrel outer membrane proteins. It is composed of the essential outer membrane protein BamA and four outer membrane associated lipoproteins BamB-E. The Yersinia enterocolitica Adhesin A (YadA) is the prototype of trimeric auotransporter adhesins (TAAs), consisting of a head, stalk and a beta-barrel membrane anchor. To investigate the role of BamA in biogenesis of TAAs, we expressed YadA in a BamA-depleted strain of Escherichia coli, which resulted in degradation of YadA. Yeast-two-hybrid experiments and immunofluorescence studies revealed that BamA and YadA interact directly and colocalize. As BamA recognizes the C-terminus of OMPs, we exchanged the nine most C-terminal amino acids of YadA. Substitution of the amino acids in position 1, 3 or 5 from the C-terminus with glycine resulted in DegP-dependent degradation of YadA. Despite degradation all YadA proteins assembled in the outer membrane. In summary we demonstrate that (i) BamA is essential for biogenesis of the TAA YadA, (ii) BamA interacts directly with YadA, (iii) the C-terminal amino acid motif of YadA is important for the BamA-dependent assembly and differs slightly compared with other OMPs, and (iv) BamA and YadA colocalize.

    KW - OUTER-MEMBRANE-PROTEIN

    KW - CONSERVED BACTERIAL PROTEIN

    KW - ESCHERICHIA-COLI

    KW - VIRULENCE PLASMID

    KW - BINDING DOMAIN

    KW - OMP85

    KW - SECRETION

    KW - PATHWAY

    KW - TRANSLOCATION

    KW - TRANSPORT

    U2 - 10.1111/j.1365-2958.2010.07377.x

    DO - 10.1111/j.1365-2958.2010.07377.x

    M3 - Article

    VL - 78

    SP - 932

    EP - 946

    JO - Molecular Microbiology

    JF - Molecular Microbiology

    SN - 0950-382X

    IS - 4

    ER -