Unusual ultrastructures of the Branchiostoma IF protein C2 containing heptads in the tail

Abstract

Branchiostoma intermediate filament (IF) protein C2 contains a long tail domain consisting of several degenerate repeats which display a heptad repeat pattern. This unique tail sequence is predicted to constitute a long coiled coil domain in C2, which is separated from the rod by a glycine-rich linker L3. The recombinant IF protein C2 shows, in electron microscopy (EM), parallel rodlike dimers of 66.7 nm decorated by a larger globule on one side and a smaller globule on the other side. In contrast, the length of the tailless C2 dimers, decorated by only one small globule, is about 26 nm shorter. These results indicate that both the rod domain and the newly predicted coiled coil segment 3 participate in the formation of a double-stranded coiled coil dimer. Moreover, the two to four C2 dimers are able to associate via their globular tail domain into multiarm oligomers, an ability not seen by the tailless C2 mutant or the other currently known protostomic and vertebrate IFs.