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Type IV-like pili formed by the type II secreton: specificity, composition, bundling, polar localization, and surface presentation of peptides
- Source :
- Journal of Bacteriology. June, 2003, Vol. 185 Issue 11-12, p3416, 13 p.
- Publication Year :
- 2003
-
Abstract
- The secreton or type II secretion machinery of gram-negative bacteria includes several type IV pilin-like proteins (the pseudopilins) that are absolutely required for secretion. We previously reported the presence of a bundled pilus composed of the pseudopilin PulG on the surface of agar-grown Escherichia coli K-12 cells expressing the Klebsiella oxytoca pullulanase (Pul) secreton genes at high levels (N. Sauvonnet, G. Vignon, A. P. Pugsley, and P. Gounon, EMBO J. 19:2221-2228, 2000). We show here that PulG is the only pseudopilin in purified pili and that the phenomenon is not restricted to the Pul secreton reconstituted in E. coli or to PulG. For example, high-level expression of the endogenous E. coli gsp secreton genes caused production of bundled pili composed of the pseudopilin GspG, and the Pul secreton was able to form pili composed of PulG-like proteins from secreton systems of other bacteria. PulG derivatives in which the C terminus was extended by the addition of eight different peptides were also assembled into pill and functioned in secretion. Three of the C-terminal peptides were shown to be exposed along the entire length of the assembled pill. Hence, the C terminus of PulG may represent a permissive site for the insertion of immunogenic epitopes or other peptide sequences. One of these PulG variants, with a six-histidine tag at its C terminus, formed nonpolar, nonbundled pili, suggesting that bundle formation and polar localization are not correlated with the ability of PulG to function in secretion. We propose that the PulG pilus is an artifactual manifestation of a periplasmic 'pseudopilus' and that cycles of pseudopilus extension and retraction within the periplasm propel pullulanase through secretin channels in the outer membrane. Abnormally long pili that extend beyond the outer membrane are produced only when pilus length control and retraction are deregulated by overproduction of the major pseudopilus subunit (PulG).
- Subjects :
- Gene expression -- Physiological aspects
Bacterial proteins -- Genetic aspects
Cytology -- Genetic aspects
Escherichia coli -- Growth
Escherichia coli -- Genetic aspects
Gram-negative bacteria -- Genetic aspects
Secretion -- Genetic aspects
Microbial populations -- Genetic aspects
Bacteriology -- Research
Company growth
Biological sciences
Subjects
Details
- Language :
- English
- ISSN :
- 00219193
- Volume :
- 185
- Issue :
- 11-12
- Database :
- Gale General OneFile
- Journal :
- Journal of Bacteriology
- Publication Type :
- Academic Journal
- Accession number :
- edsgcl.107217841