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  • Title: The role of lamin LIII in nuclear assembly and DNA replication, in cell-free extracts of Xenopus eggs.
    Author: Meier J, Campbell KH, Ford CC, Stick R, Hutchison CJ.
    Journal: J Cell Sci; 1991 Mar; 98 ( Pt 3)():271-9. PubMed ID: 2055960.
    Abstract:
    Xenopus egg extracts, which support nuclear assembly and DNA replication, were functionally depleted of lamin LIII by inoculating them with monoclonal anti-lamin antibodies. Phase-contrast microscopy and electron-microscopy studies indicated that lamin-depleted extracts supported efficient chromatin decondensation, and assembly of double membrane structures and nuclear pores on demembranated sperm heads. Immunofluorescence microscopy suggests that lamin-antibody complexes are transported across the nuclear membrane but do not assemble into a lamina. These findings were confirmed by immunoblotting analysis of isolated nuclei. Metabolic labelling studies with either biotin-11-dUTP or [32P]dCTP, revealed that nuclei lacking a lamina were unable to initiate DNA replication and that, although such nuclei could import proteins required for DNA replication (e.g. PCNA), these proteins were apparently not organized into replicon clusters.
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