Temperature-sensitive intermediate filament assembly. Alternative structures of Xenopus laevis vimentin in vitro and in vivo.
In assembly assays of intermediate filaments (IFs) from vimentin of the amphibian species Xenopus laevis we have observed the formation of so far unknown structures at temperatures above 28 degrees C. Upon assembly in vitro at temperatures above 34 degrees C massive aggregates, partly with a protofilamentous substructure, were found and their formation correlated with drastically reduced end-viscosity. Large spheroidal, dense aggregates with a complex suborganization were also seen to form at 37 degrees C in the cytoplasm of living mammalian cells devoid of endogenous vimentin upon transfection with cDNA encoding the amphibian vimentin, and this was also true for vimentin forced to accumulate in the nucleoplasm by the introduction of a "nuclear localization signal". Upon shift from the non-permissive (37 degrees C) to the permissive (28 degrees C) temperature, such aggregates of non-IF vimentin structures gradually disappeared and a normal-looking IF meshwork formed. The results, which are discussed in relation to other structures assembled by IF proteins, indicate a marked thermosensitivity in the amino acid sequence of the vimentin which seems to have been reduced during evolution of warm-blooded animals. They further show that members of the multigene gene family of IF proteins can occur in structures totally different from IFs.
Herrmann
H
H
Division of Cell Biology, German Cancer Research Center, Heidelberg.
Eckelt
A
A
Brettel
M
M
Grund
C
C
Franke
W W
WW
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D066298:In Vitro Techniques; D007382:Intermediate Filaments / Q000648:ultrastructure*; D008854:Microscopy, Electron; D011994:Recombinant Proteins; D013045:Species Specificity; D013696:Temperature; D014162:Transfection; D014746:Vimentin / Q000502:physiology*; D014783:Viscosity; D014982:Xenopus laevis / Q000502:physiology*
D011994:Recombinant Proteins; D014746:Vimentin