Pilus

Escherichia coli cell with bacterial fimbriae (pili). EM ~10,000 fold magnification. From: Bacterial Fimbriae Designed to Stay with the Flow Gross L PLoS Biology Vol. 4, No. 9, e314 doi:10.1371/journal.pbio.0040314 Creative Commons Attribution License (CCAL).

A pilus (Latin for 'hair'; plural : pili) is a hairlike protein structure on the surface of a cell, especially Gram-negative bacteria.

A pilus is typically 7 nm in diameter , and several hundred pili can extend from the surface of a bacterial cell (see image to right).

A fimbrium (Latin meaning fringe; plural: fimbriae) is a short pilus that is used to attach the cell to a surface. Fimbriae are either located at the poles of a cell, or are evenly spread over its entire surface. In some cases, mutant pathogenic bacteria that lack fimbriae cannot adhere to their usual target host cell surfaces, and thus cannot cause disease.

The terms pili and fimbriae are often used interchangeably. Numerous different types of pili have been characterized, and various forms of these appendages are involved in diverse activities of bacteria. These include bacterial cell aggregation, adhesion to surfaces of host cells such as, in the case of gut bacteria, the linings of the intestine, adhesion to other microbial cells in biofilms, gene and protein injection into other cells, DNA uptake by naturally transformable bacteria, and virulence attributes of pathogenic bacteria.

Pili are distinctly different from bacterial flagella; unlike flagella they grow from the inside of the cell outward, and not from the tip of the fiber.

Bacterial fimbriae can contain lectin proteins generally at their tips. These lectins are necessary to adhere to target cells because they can recognize oligosaccharide units such as mannose on the surface of these target cells. However, all types of pili are primarily composed of oligomeric pilin proteins.

Some forms of pili are encoded by plasmids, for instance fertility factor F encodes F-pili (sex-pili) which are present at only 2 to 4 per cell. F-pili are involved in bacterial conjugation and connect the bacterium to another bacterium and build a bridge between the cytoplasms of the cells which becomes the channel for one way transfer of a single-strand of DNA and certain protein molecules. This transfer enables the mobilization of plasmids between bacteria. An exchanged plasmid can add new functions to a bacterium, e.g., an antibiotic resistance.

Some bacterial viruses (bacteriophages) attach to receptors on sex pili at the start of their reproductive cycle.

Type I pili

Type I pili are widely distributed on enteric bacteria and their structure and assembly is well characterized. They are composed of several different protein components including FimH adhesin that is part of the short thin fibrillar pilus tip, and FimA which makes up the thicker 7 nm thick long rod of the pilus. FimA binds to mannose sugars present on a variety of different host cell surface structures and can be considered to be a lectin .

The assembly of the Type I pilus occurs in the periplasm and involves the "chaperone/usher" protein folding and delivery system.

Sex pili

Schematic drawing of bacterial conjugation. 1- Donor cell produces pilus. 2- Pilus attaches to recipient cell, brings the two cells together. 3- The mobile plasmid is nicked and a single strand of DNA is then transferred to the recipent cell. 4- Both cells recircularize their plasmids, synthesize second strands, and reproduce pili; both cells are now viable donors.

The pili encoded by F plasmid in Escherichia coli are involved in formation of cell aggregates typically 2 to 13 cells in number. Numerous genes on the F-plasmid are involved in pilus formation including genes traA, E K B V W U F and G .

Adhesion of bacterial cells has an important survival role in their survival as micro-colonies - called biofilms - on solid surfaces in the natural environment, and F-pili determine the final shapes of the structures seen in mature surface biofilms formed by Eschericha colibacteria, as mutants affected in the plasmid specified F-pili form a biofilm of a different structure.

The pilus allows for the transfer of bacterial DNA from the F-piliated (donor) bacteria to the recipient bacteria. Through this mechanism of gene transfer, advantageous genetic traits can be disseminated amongst a population of bacteria.

Not all bacteria have the ability to create sex pili, however sex pili can form mating channels between bacteria of different species, and even between bacteria and eukaryotic cells such as those of yeasts and plants. The VirB protein of croen-gall bacterium Agrobacterium tumefaciens is a pilin like protein similar to TraA of E. coli F-plasmid that is involved in the injection of T-DNA into plant cells by the crown-gall bacterium.

Type 4 pili

Type 4 pili are thin, flexible , 6- to 7-nm fibers displayed by a wide variety of gram-negative bacterial species and members of this class are recognisable because of amino-acid sequence similarities. They have a distinctive method of assembly involving proteolytic processing and N-methylation of the pilin precursor, and they are not implicated in conjugation. Components of the pilus are also involved in natural DNA uptake (transformation) in Neisseria bacteria. Type 4 pili are primarily involved in adhesion to surfaces and as virulence determinants in pathogenic bacteria. In some species they cause twitching motility, for instance in Pseudomonas aeruginosa, Myxococcus and Neisseria bacteria .

The pathogenic diplococcus coccus Neisseria gonorrhoeae has been shown to move by extension of Type 4 pili so that external termini of the pili adhere to solid substrate, and subsequent pili retraction pulls the bacteria forward, not unlike a grappling hook .

Pili of pathogens

Type 4 pili are produced by the enteric pathogenic bacteria Vibrio cholerae, enteropathogenic Escherichia coli, and enterotoxigenic E. coli. Bundle forming pili (BfpA) of enteropathogenic Escherichia coli are Type IV pili that are necessary for adhesion to adhere to epithelial cells. Type 4 pili are virulence factors of Neisseria meningitidis and N. gonorrhoeae bacteria. Many bacteria producing Type 4 pili also produce other adhesion factors .

Illustrations of these pilus-target interactions, and of the remarkable design features of these powerful bacterial grappling devices, are found in the PLoS Biology article that follows.

Bacterial fimbriae designed to stay with the flow

Liza Gross

The human digestive system houses a diverse colony of beneficial bacteria, but one species—E. coli—can wreak havoc when it colonizes mucous membranes that normally exist unmolested (for example, in the urinary tract). To latch on to cells and establish infection, E. coli uses fimbriae—long, hairlike organelles that project from the bacterium’s surface. Fimbriae consist of interlinking subunits of a single protein called pilin that forms a rigid, coiled helix-shaped rod. Sticky proteins called adhesins cap the tip of the rod and bind to carbohydrate receptors on their host, thus securing bacteria on the host cells as extracellular fluids swirl around them.

A previous study led by Evgeni Sokurenko and Viola Vogel investigated the most common type of E. coli fimbriae. The sticky protein at the tip of these fimbriae is called FimH and binds to a carbohydrate called mannose. They showed that powerful drag forces created by the extracellular fluids don’t carry the bound bacteria away, as one might expect, but instead strengthen their adhesion to their host. The researchers attributed this increased binding to a biphasic “catch bond” mechanism whereby increased drag forces cause the FimH at the tip of the fimbria to switch from a form that binds mannose weakly to a form that binds strongly. Because of this, the bacteria bind best at an optimal force that is high enough to switch FimH to strong binding but not so high that it breaks the strong FimH–mannose bond.

And now, in a new study, the same group of researchers (including first author Manu Forero) set out to determine whether the coiled rod structure of fimbriae affects how the sticky FimH at the tip binds. It had been assumed that fimbrial rods play a largely static structural role, either by extending the tip adhesins’ reach or by resisting electrostatic repulsive forces between bacteria and cell surfaces. But Forero et al. show that the rods function more dynamically, using their mechanical properties to help stabilize the FimH–mannose bond against a turbulent background.

Fimbriae-mediated adhesion was investigated with an atomic force microscope, which uses a cantilever to apply (and measure) forces between its tip and the sample under investigation. Forero et al. outfitted the cantilever tip with mannose, and then used this to touch a fimbriated E. coli cell that was affixed to a glass surface. After mannose bound to the fimbrial FimH, the cantilever retreated from the bacterium at a constant velocity. The researchers determined that, instead of the FimH–mannose bond breaking, the fimbriae stretched out far beyond their original length.

One reason that fimbriae extend could be that the individual pilin subunits of the fimbrial rod are uncoiling. The researchers tested this hypothesis by applying a constant force between the cantilever and fimbria—under which fimbrial length changes slowly. They observed that stepwise jumps in distance corresponded to the expected length of individual subunits unwinding from the coiled shaft one at a time. Thus, the researchers concluded, fimbrial elongation proceeds as subunits uncoil one after another. This was also supported by a mathematical model developed by the researchers to quantify the biophysical forces governing the dynamics of fimbrial uncoiling.

Forero et al. also detected that, after uncoiling at increasing force, the stretched fimbriae re-coil if the pulling force drops. Importantly, while fimbrial uncoiling under high force decreases the tension within the rod, re-coiling under low force increases the tension. Thus, the tensile force within the rod stays within some intermediate level when fimbrial length is stable.

The researchers found that the intermediate force range corresponds to the force level where the FimH–mannose bonds last longest. Lower, coiling forces are too weak to switch bonds to a long-lived state before breaking, and higher, uncoiling forces exceed the catch-bond threshold, shortening the life of the bond. Because E. coli living in the gut or other mucosal surfaces experience constantly changing flow rates and forces, these adjustments should enhance fimbrial attachment under a diverse range of fluid conditions. The correspondence of forces suggests that the mechanical properties of the fimbrial rod and the FimH–mannose complex co-evolved to optimize adhesive stability in fluids.

Citation: Gross L (2006) Bacterial Fimbriae Designed to Stay with the Flow. PLoS Biol 4(9): e314 DOI: 10.1371/journal.pbio.0040314

Published: August 29, 2006

Copyright: © 2006 Public Library of Science. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

References

Further reading

• CH Jones, JS Pinkner, R Roth, J Heuser, AV Nicholes, SN Abraham and SJ Hultgren (1995) FimH Adhesin of Type 1 pili is Assembled into a Fibrillar tip Structure in the Enterobacteriaceae Proceedings of the National Academy of Sciences, Vol 92, pages 2081-2085

See also

• Flagellum