Disassembly of the Drosophila nuclear lamina in a homologous cell-free system.
Stage 14 Drosophila oocytes are arrested in first meiotic metaphase. A cell-free extract of these oocytes catalyzes apparent disassembly of purified Drosophila nuclei as well as of nuclear lamin polymers formed in vitro from isolated interphase lamins. Biochemically, the oocyte extract catalyzes lamin solubilization and phosphorylation as well as characteristic changes in one- and two-dimensional gel mobility. A previously unidentified soluble lamin isoform is easily seen after in vitro disassembly. This isoform is detectable but present only in very small quantities in vivo and is apparently derived specifically from one of the two interphase lamin isoforms. Cell-free nuclear lamina disassembly is ATP-dependent and addition of calcium to extracts blocks disassembly as judged both morphologically and biochemically. This system will allow enzymological characterization of cell-free lamina disassembly as well as molecular analysis of specific Drosophila mutants.
Maus
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N
Graduate Program in Genetics, State University of New York at Stony Brook 11794-8651, USA.
Stuurman
N
N
Fisher
P A
PA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002457:Cell Extracts / Q000494:pharmacology; D002474:Cell-Free System; D002478:Cells, Cultured; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism* / Q000648:ultrastructure; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D034882:Lamins; D008540:Meiosis; D009685:Nuclear Envelope / Q000187:drug effects / Q000378:metabolism*; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009865:Oocytes / Q000737:chemistry*; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects
D002457:Cell Extracts; D034882:Lamins; D009687:Nuclear Proteins; D000255:Adenosine Triphosphate; D002118:Calcium