In order to view the three-dimensional arrangement of these unique structures within intact bacteria, the researchers collaborated with the laboratory of Grant Jensen, professor of biology and HHMI investigator at Caltech. Utilizing a technique called electron cryotomography, the researchers flash-froze the bacterial cells at very low temperatures. This allowed them to view the cells and their internal structures in their natural, "near-native" states.
Using this visualization technique, Martin Pilhofer, a postdoctoral scholar in Jensen's lab and the paper's other first author, discovered something unique about the phage tail-like structures within P. luteoviolacea; instead of existing as individual appendages, the structures were linked together to create a spiny array. "In these arrays, about 100 tails are stuck together in a hexagonal lattice to form a complex with a porcupine-like appearance," Shikuma says. "They're all facing outward, poised to fire," he adds. "We believe this is the first observation of arrays of phage tail-like structures."
Initially, the array is compacted within each bacterium; however, the cells eventually burstkilling the microbesand the array unfolds. The researchers hypothesize that, at this point, the individual spines of the array fire outward into the tubeworm larva. Following this assault, the larvae begin their developmental transition to adulthood.
"It was a tremendous surprise that the agent that drives metamorphosis is such an elaborate, well-organized injection machine," says coauthor Jensen. "Who would have guessed that the signal is delivered by an apparatus that is almost as large as the bacterial cell itself? It is simply a marvelous structure, synthesized in a 'loaded' but tightly collapsed state within the cell, which then expands like an umbrella, opening up into a much larger web of syringes that are ready to inject," he says.
Although the study confirms that the phag
|Contact: Deborah Williams-Hedges|
California Institute of Technology