Images reveal how rotavirus - leading cause of diarrhea - enters cells

kes," each consisting of a cluster of VP4 molecules. Dormitzer and colleagues trimmed the VP4 protein down to two rocklike sub-components that make up the spike's "head" and "body." They crystallized these pieces and used X-ray diffraction to determine their three-dimensional structures, precise down to the atom. Comparison of the crystal structures to electron microscopy images, obtained by colleagues at the Baylor College of Medicine, suggests that VP4 undergoes two consecutive shape changes that allow it to breach the membrane of the cell it's trying to infect.

"This protein goes through some extraordinary gymnastics that are almost certainly related to its function," says Dormitzer.

First, when rotavirus arrives in the intestine, digestive enzymes cause two of the three VP4 molecules in each cluster to form a rigid spike, priming the virus to attack the cell and positioning the spike "head" to bind to the surface of the target cell. Then, in a second rearrangement, the spikes fold back, and VP4 takes on a folded-umbrella structure with three "panels." Dormitzer and colleagues speculate that this folding motion causes the spike's "body" component to break a hole in the cell membrane, allowing the virus to enter.

The key finding for vaccine development is that the "head" and "body" portions of the VP4 protein contain many of the targets that the immune system recognizes when it attacks the virus and protects against infection.

"The work is a clear example of the way in which structural studies can contribute to new good ideas about strategies for vaccines," says Senior Investigator Stephen Harrison, Ph.D., a Howard Hughes Medical Institute investigator and chief of CHB's Laboratory of Molecular Medicine.

Vaccines made from live viruses (such as RotaShield) or killed viruses can present safety concerns and be unstable without refrigeration. In contrast, the "head" of the VP4 protein is very stable at room temperature

Contact: Mary-Ellen Shay
Children's Hospital Boston

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