Cellular localization of Drosophila 83-kilodalton heat shock protein in normal, heat-shocked, and recovering cultured cells with a specific antibody.
To gain insight on the possible functions of heat shock proteins (hsp's) in Drosophila, we have purified the 83-kilodalton hsp (hsp 83) from cultured cells and studied its intracellular localization by immunofluorescence in normal, heat-shocked, and recovering cells. The specificity of the antibody was assessed by one- and two-dimensional gel immunoblotting and by partial proteolytic digestion. The anti-hsp 83 antibody does not show any significant cross-reactivity with hsp's of different avian or mammalian cell lines, but cross-reacts with hsp's of similar molecular masses in other dipteran insects. The partial proteolytic peptide maps of Drosophila hsp 83 differ from those of mouse hsp 89 and chicken hsp 84. Immunoblotting of Drosophila Kc cells heat shocked at different temperatures indicates a maximal expression of hsp 83 at 33 degrees C. By immunofluorescence, hsp 83 is shown to have a strictly cytoplasmic localization. In unstressed cells, it is distributed in the entire cytoplasm with a slight enrichment in the perinuclear region. After heat shock, it seems to concentrate at the cell periphery close to the plasma membrane and it gradually redistributes to the whole cytoplasm during cellular recovery at normal temperatures.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004331:Drosophila melanogaster / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006360:Heat-Shock Proteins / Q000276:immunology / Q000378:metabolism*; D006358:Hot Temperature; D013045:Species Specificity
D000906:Antibodies; D000939:Epitopes; D006360:Heat-Shock Proteins