Nuclei that lack a lamina accumulate karyophilic proteins and assemble a nuclear matrix.
Xenopus egg extracts, which support nuclear assembly and DNA replication in vitro, were physically depleted of lamin B3 using monoclonal antibodies linked to magnetic beads. Depleted extracts were still able to support nuclear envelope assembly around demembranated sperm heads but the resulting pronuclei lacked a lamina and were unable to initiate semiconservative DNA replication or to assemble replicases, confirming previous data. Immunoblotting analysis of isolated nuclei and nuclear matrix fractions indicated that lamin-depleted nuclei still accumulated nucleoporins and PCNA. Furthermore, the rate of PCNA uptake was identical in lamin-depleted and control nuclei. However, neither the nucleoporins nor the PCNA was associated with nuclear matrix fractions. The major protein components of sperm pronuclear matrix fractions were characterized by two-dimensional gel electrophoresis. Of these proteins only three out of 22 species, other than the lamins, were significantly reduced in lamin-depleted nuclei, indicating that these nuclei do assemble a nuclear matrix.
Jenkins
H
H
Department of Biological Sciences, The University, Dundee, UK.
Hölman
T
T
Lyon
C
C
Lane
B
B
Stick
R
R
Hutchison
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D004261:DNA Replication; D005260:Female; D018189:Immunomagnetic Separation; D007381:Intermediate Filament Proteins; D034921:Lamin Type B; D034882:Lamins; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009685:Nuclear Envelope / Q000378:metabolism*; D015530:Nuclear Matrix / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D009865:Oocytes / Q000737:chemistry / Q000648:ultrastructure; D018809:Proliferating Cell Nuclear Antigen; D013077:Sperm Head / Q000502:physiology / Q000648:ultrastructure; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins; D034921:Lamin Type B; D034882:Lamins; D008562:Membrane Glycoproteins; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; C049312:nuclear pore protein p62