1887

Abstract

thin aggregative fimbriae (Tafi; curli) are important in pathogenesis and biofilm formation; however, less is known of their structure and morphogenesis. In the Tafi operon, the transcription and role of have been elusive. In this study, transcripts were detected using a sensitive reverse transcriptase technique. Native AgfC was not detected using polyclonal antibodies generated against purified hexahistidine-tagged AgfC; however, expression revealed that AgfC was localized to the periplasm as a mature form. An isogenic Δ mutant displayed an abundance of 20 nm fibres, in addition to native Tafi (5–7 nm), and had an increase in cell surface hydrophobicity. Purified 20 nm fibres were depolymerized under exceptionally stringent conditions to release what proved to be AgfA subunits. This revealed that the 20 nm fibres represented a different form of Tafi. The role of AgfC in Tafi assembly was investigated further using an antibody-capture assay of isogenic Δ mutants. A soluble antibody-accessible form of AgfA was captured in wild-type (), Δ and Δ strains, in support of the extracellular nucleation–precipitation pathway of Tafi assembly, but not in Δ or Δ mutants. This indicates that AgfC and AgfE are important for AgfA extracellular assembly, facilitating the synthesis of Tafi.

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2007-04-01
2020-10-01
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