title: Bypassing the need for subcellular localization of a polysaccharide export-anchor complex by overexpressing its protein subunits.
abstract: Subcellular protein localization is thought to promote protein-protein interaction by increasing the effective concentration and enabling spatial co-ordination and proper segregation of proteins. We found that protein overexpression allowed the assembly of a productive polysaccharide biosynthesis-export-anchoring complex in the absence of polar localization in Caulobacter crescentus. Polar localization of the holdfast export protein, HfsD, depends on the presence of the other export proteins, HfsA and HfsB, and on the polar scaffold protein PodJ. The holdfast deficiency of hfsB and podJ mutants is suppressed by the overexpression of export proteins. Restored holdfasts are randomly positioned and colocalize with a holdfast anchor protein in these strains, indicating that functional complexes can form at non-polar sites. Therefore, overexpression of export proteins surpasses a concentration threshold necessary for holdfast synthesis. Restoration of holdfast synthesis at non-polar sites reduces surface adhesion, consistent with the need to spatially co-ordinate the holdfast synthesis machinery with the flagellum and pili. These strains lack the cell-specific segregation of the holdfast, resulting in the presence of holdfasts in motile daughter cells. Our results highlight the fact that multiple facets of subcellular localization can be coupled to improve the phenotypic outcome of a protein assembly.
authors.affiliation: Department of Biology, Indiana University, Bloomington, IN 47405, USA.
authors.forename: June
authors.initials: J
authors.lastname: Javens
authors.forename: Zhe
authors.initials: Z
authors.lastname: Wan
authors.forename: Gail G
authors.initials: GG
authors.lastname: Hardy
authors.forename: Yves V
authors.initials: YV
authors.lastname: Brun
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016935:Caulobacter crescentus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D016764:Cell Polarity; D010861:Fimbriae, Bacterial / Q000235:genetics / Q000378:metabolism; D005407:Flagella / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011134:Polysaccharides / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D011134:Polysaccharides; D021122:Protein Subunits; C120058:podj protein, Caulobacter crescentus