Analysis of cytoplasmic 19 S ring-type particles in Drosophila which contain hsp 23 at normal growth temperature.
Cytoplasmic 19 S particles were isolated from postpolysomal supernatants of 25 degrees C Drosophila embryos and culture cells. The particles were purified by salt extraction and sucrose gradient centrifugation. Electron microscopic investigation showed that the 19 S particles possess a ring-shaped morphology with an outer diameter of 12 nm and a hollow core of 3 nm. Biochemically the particles are characterized by a group of 16 polypeptides within the molecular weight range of 35 to 23 kDa, and small RNA molecules in the size range of 200 to 60 nucleotides. The RNP character of the particles is also shown by their buoyant density in Cs2SO4 of rho = 1.29 g/cm3 and their susceptibility to uv crosslinking and density in CsCl of rho = 1.38 g/cm3. Antibodies were raised against the proteins of the 19 S particles isolated from 25 degrees C cells and tested by immunoblotting after one- and two-dimensional gel-electrophoresis. Two of the antibodies raised cross react with the small heat-shock proteins hsp 28/27 and hsp 23. Comparative protease V8 cleavage of hsp 23 and the 23-kDa particle protein demonstrates that these two proteins are identical and that the small hsp of Drosophila must be a genuine part of the 19 S cytoplasmic ring-shaped complexes at normal growth temperature. The data support the idea of a general developmental role of some of the so-called heat-shock proteins.
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D000818:Animals; D002499:Centrifugation, Density Gradient; D003593:Cytoplasm / Q000032:analysis; D004331:Drosophila melanogaster / Q000032:analysis* / Q000254:growth & development; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000032:analysis*; D007163:Immunosorbent Techniques; D002479:Inclusion Bodies / Q000032:analysis*; D008854:Microscopy, Electron; D008970:Molecular Weight; D012313:RNA / Q000032:analysis; D013367:Submitochondrial Particles / Q000032:analysis; D013696:Temperature
D006360:Heat-Shock Proteins; D012313:RNA