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    Agrobacterium VirB10 domain requirements for type IV secretion and T pilus biogenesis

    Jakubowski, S.J. and Kerr, J.E. and Garza, I. and Krishnamoorthy, V. and Bayliss, R. and Waksman, Gabriel and Christie, P.J. (2009) Agrobacterium VirB10 domain requirements for type IV secretion and T pilus biogenesis. Molecular Microbiology 71 (3), pp. 779-794. ISSN 0950-382X.

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    Agrobacterium tumefaciens VirB10 couples inner membrane (IM) ATP energy consumption to substrate transfer through the VirB/D4 type IV secretion (T4S) channel and also mediates biogenesis of the virB-encoded T pilus. Here, we determined the functional importance of VirB10 domains denoted as the: (i) N-terminal cytoplasmic region, (ii) transmembrane (TM) α-helix, (iii) proline-rich region (PRR) and (iv) C-terminal β-barrel domain. Mutations conferring a transfer- and pilus-minus (Tra-, Pil-) phenotype included PRR deletion and β-barrel substitution mutations that prevented VirB10 interaction with the outer membrane (OM) VirB7–VirB9 channel complex. Mutations permissive for substrate transfer but blocking pilus production (Tra+, Pil-) included a cytoplasmic domain deletion and TM domain insertion mutations. Another class of Tra+ mutations also selectively disrupted pilus biogenesis but caused release of pilin monomers to the milieu; these mutations included deletions of α-helical projections extending from the β-barrel domain. Our findings, together with results of Cys accessibility studies, indicate that VirB10 stably integrates into the IM, extends via its PRR across the periplasm, and interacts via its β-barrel domain with the VirB7–VirB9 channel complex. The data further support a model that distinct domains of VirB10 regulate formation of the secretion channel or the T pilus.


    Item Type: Article
    School: Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences
    Research Centres and Institutes: Structural Molecular Biology, Institute of (ISMB)
    Depositing User: Administrator
    Date Deposited: 04 Aug 2010 14:09
    Last Modified: 02 Aug 2023 16:49


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