title: Nucleator function of CsgB for the assembly of adhesive surface organelles in Escherichia coli.
abstract: Curli are surface organelles in Escherichia coli that assemble outside the bacterium through the precipitation of secreted soluble CsgA monomers, requiring the CsgB nucleator protein. Using immunoelectron microscopy and immunoblotting assays, CsgB is shown to be located on the bacterial surface and also as a minor component of wild-type curli. CsgB lacking its 20 N-terminal residues when fused to maltose-binding protein (MBP) can still trigger polymerization of CsgA monomers in vivo. However, the resulting surface organelles are only formed at one of the two bacterial poles and are morphologically distinct from wild-type curli. These Bfco organelles (CsgB-Free Curli-related Organelles) are highly regular structures reacting with anti-CsgA, but not anti-CsgB antibodies. The CsgB of the active MBP-CsgBII fusion is surface exposed but, unlike the native CsgB in wild-type curli, is not detectable in the Bfco organelles. Overexpression of csgB within a csgA mutant results in the formation of short CsgB polymers on the cell surface. It is suggested that in wild-type bacteria, both CsgA and CsgB are secreted proteins. Interaction between CsgA and CsgB triggers wild-type curli formation, resulting in CsgA-CsgB heteropolymers, while surface-anchored CsgB in MBP-CsgBII triggers morphologically distinct, CsgB-free/CsgA Bfco organelles. In the absence of CsgA, CsgB can self-assemble into polymers.
authors.affiliation: Microbiology and Tumorbiology Center (MTC), Karolinska Institute, S-171 77 Stockholm, Sweden.
authors.forename: Z
authors.initials: Z
authors.lastname: Bian
authors.forename: S
authors.initials: S
authors.lastname: Normark
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D015153:Blotting, Western; D004926:Escherichia coli / Q000378:metabolism* / Q000648:ultrastructure; D029968:Escherichia coli Proteins; D005353:Fibronectins / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D015388:Organelles / Q000502:physiology*; D011108:Polymers / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; C109182:CsgB protein, E coli; D029968:Escherichia coli Proteins; D005353:Fibronectins; D011108:Polymers; D011993:Recombinant Fusion Proteins; C489688:csgA protein, E coli; C077481:Crl protein, Bacteria