A conserved region in the tail domain of vimentin is involved in its assembly into intermediate filaments.
Although the head and rod domains of intermediate filament (IF) proteins are known to play significant roles in filament assembly, the role of the tail domain in this function is unclear and the available information supports contradictory conclusions. We examined this question by comparing transfection of the same cDNA constructs, encoding vimentins with modified tail domains, into cell lines that do and do not contain endogenous IF proteins. By this approach, we were able to distinguish between the ability of a mutant IF protein to initiate assembly de novo, from that of incorporating into existing filament networks. Vimentins with modifications at or near a highly conserved tripeptide, arg-asp-gly (RDG), of the tail domain incorporated into existing IF networks in vimentin-expressing (vim+) cells, but were assembly-incompetent in cells that did not express IF proteins (vim-). The failure of the RDG mutant vimentins to assemble into filament arrays in vim- cells was reversible by re-introducing a wild-type vimentin cDNA, whereupon both wild-type and mutant vimentins coassembled into one and the same IF network. We conclude that the function of the tail domain of type III IF proteins, and possibly of keratins K8 and K18, in IF assembly is distinct from those of other domains; a region encompassing the RDG tripeptide appears to be important in the assembly process.
Department of Anatomy and Cell Biology, University of Cincinnati College of Medicine.
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Makarova
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Carpenter
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Khan
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D000310:Adrenal Gland Neoplasms / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D001120:Arginine / Q000032:analysis; D001224:Aspartic Acid / Q000032:analysis; D001483:Base Sequence; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D005354:Fibrosarcoma / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D005998:Glycine / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000378:metabolism*; D008969:Molecular Sequence Data; D010641:Phenotype; D014162:Transfection; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*
D004273:DNA, Neoplasm; D014746:Vimentin; D001224:Aspartic Acid; D001120:Arginine; D005998:Glycine