Selective digestion of nuclear envelopes from Xenopus oocyte germinal vesicles: possible structural role for the nuclear lamina.
We have used enzymic digestion as a structural probe to investigate components of the nuclear envelope of germinal vesicles from Xenopus oocytes. Previous studies have shown that these envelopes are composed of a double membrane in which nuclear pore complexes are embedded. The nuclear pore complexes are linked to a fibrous lamina that underlies the nucleoplasmic face of the envelope. The pores are also linked by pore-connecting fibrils that attach near their cytoplasmic face. Xenopus oocyte nuclear envelopes were remarkably resistant to extraction with salt solutions and, even after treatment with 1 M NaCl or 3 M MgCl2, pores, lamina and pore-connecting fibrils remained intact. However, mild proteolysis with trypsin selectively removed the lamina fibres from Triton-extracted nuclear envelopes to leave only the pore complexes and connecting fibrils. This observation confirmed that the pore-connecting fibrils were different from the lamina fibres and were probably constructed from different proteins. Trypsin digestion followed by Triton treatment resulted in the complete disintegration of the nuclear envelope, providing direct evidence for a structural role for the lamina in maintaining envelope integrity. Digestion with ribonuclease did not produce any marked change in the structure of Triton-extracted nuclear envelopes, indicating that probably neither the pore-connecting fibrils nor the cytoplasmic granules on the pore complexes contained a substantial proportion of RNA that was vital for their structural integrity.
Whytock
S
S
MRC Laboratory of Molecular Biology, UK.
Moir
R D
RD
Stewart
M
M
D000818:Animals; D003101:Collodion; D003902:Detergents; D034882:Lamins; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009685:Nuclear Envelope / Q000502:physiology* / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000502:physiology*; D017830:Octoxynol; D009865:Oocytes / Q000648:ultrastructure; D010447:Peptide Hydrolases; D011092:Polyethylene Glycols; D012259:Ribonuclease, Pancreatic; D013194:Staining and Labeling; D014982:Xenopus laevis
D003902:Detergents; D034882:Lamins; D009687:Nuclear Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; D003101:Collodion; D012259:Ribonuclease, Pancreatic; D010447:Peptide Hydrolases