Characterization of an Intermediate Filament Protein from the Platyhelminth, Dugesia japonica.

Akiko Yamamoto, Ken-Ichiro Matsunaga, Toyoaki Anai, Hitoshi Kawano, Toshihisa Ueda, Toshihiko Matsumoto, Shoji Ando

Protein and peptide letters 2020

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Intermediate Filaments (IFs) are major constituents of the cytoskeletal systems in animal cells. To gain insights into the structure-function relationship of invertebrate cytoplasmic IF proteins, we characterized an IF protein from the platyhelminth, Dugesia japonica, termed Dif-1. cDNA cloning, in situ hybridization, immunohistochemical analysis, and IF assembly experiments in vitro using recombinant Dif-1, were performed for protein characterization. The structure deduced from the cDNA sequence showed that Djf-1 comprises 568 amino acids and has a tripartite domain structure (N-terminal head, central rod, and C-terminal tail) that is characteristic of IF proteins. Similar to nuclear IF lamins, Djf-1 contains an extra 42 residues in the coil 1b subdomain of the rod domain that is absent from vertebrate cytoplasmic IF proteins and a nuclear lamin-homology segment of approximately 105 residues in the tail domain; however, it contains no nuclear localization signal. In situ hybridization analysis showed that Djf-1 mRNA is specifically expressed in cells located within the marginal region encircling the worm body. Immunohistochemical analysis showed that Djf-1 protein forms cytoplasmic IFs located close to the microvilli of the cells. In vitro IF assembly experiments using recombinant proteins showed that Djf-1 alone polymerizes into IFs. Deletion of the extra 42 residues in the coil 1b subdomain resulted in the failure of IF formation. Together with data from other histological studies, our results suggest that Djf- 1 is expressed specifically in anchor cells within the glandular adhesive organs of the worm and that Djf-1 IFs may play a role in protecting the cells from mechanical stress. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Citation

Akiko Yamamoto, Ken-Ichiro Matsunaga, Toyoaki Anai, Hitoshi Kawano, Toshihisa Ueda, Toshihiko Matsumoto, Shoji Ando. Characterization of an Intermediate Filament Protein from the Platyhelminth, Dugesia japonica. Protein and peptide letters. 2020;27(5):432-446