Independent expression and assembly properties of heterologous lamins A and C in murine embryonal carcinomas.
The majority of cells derived from adult mammalian tissues contain three major species of nuclear lamin proteins, A, B and C. In contrast, embryonic cells including undifferentiated murine embryonal carcinomas, contain only B-type lamins, A and C appearing only after differentiation. Human lamins A or C have been introduced by transfection into undifferentiated P19 embryonal carcinomas. Twenty-four hours after transfection, both of these proteins were found to independently associate with the nuclear envelope as judged by immunofluorescence microscopy and at the same time were associated with a salt-resistant structure having solubility properties similar to those of the nuclear lamina. Biosynthetic experiments indicated that heterologous lamin A underwent processing to its mature molecular weight, an event which in adult type cells occurs after assembly into the lamina. Observations on mitotic cells demonstrate that either of the two human lamins will, independent of the other, become dispersed throughout the cytoplasm during prophase and subsequently reassemble at the nuclear periphery during telophase. Nuclear lamins A and C are not, however, equivalent in their abilities to incorporate into the nuclear lamina in these cells. Experiments involving cells arrested in S phase using thymidine suggest that lamin C, but not lamin A, requires progression through the cell cycle and probably mitosis for assembly into the nuclear lamina of P19 EC cells.
Horton
H
H
Department of Cellular and Molecular Physiology, Harvard Medical School, Boston, MA 02115.
McMorrow
I
I
Burke
B
B
D000818:Animals; D002478:Cells, Cultured; D054278:Embryonal Carcinoma Stem Cells; D006367:HeLa Cells; D006801:Humans; D034904:Lamin Type A; D034882:Lamins; D008938:Mitosis / Q000502:physiology; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D014411:Neoplastic Stem Cells / Q000378:metabolism; D009685:Nuclear Envelope / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured
D034904:Lamin Type A; D034882:Lamins; D009687:Nuclear Proteins; C440308:lamin C