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.

[1]  D. Burns,et al.  Identification of protein kinase C (PKC) phosphorylation sites on human lamin B. Potential role of PKC in nuclear lamina structural dynamics. , 1993, The Journal of biological chemistry.

[2]  P. Traub,et al.  Identification of novel phosphorylation sites in murine A-type lamins. , 1993, European journal of biochemistry.

[3]  S. Molloy,et al.  p34cdc2 kinase-mediated release of lamins from nuclear ghosts is inhibited by cAMP-dependent protein kinase. , 1992, Experimental cell research.

[4]  J. Watson,et al.  Isopentenoid synthesis in embryonic Drosophila cells: prenylated protein profile and prenyl group usage. , 1992, Archives of biochemistry and biophysics.

[5]  M. Peter,et al.  Mitogen-activated protein kinases phosphorylate nuclear lamins and display sequence specificity overlapping that of mitotic protein kinase p34cdc2. , 1992, European journal of biochemistry.

[6]  E. Nigg.  Assembly-disassembly of the nuclear lamina , 1992, Current Biology.

[7]  M. Peter,et al.  Disassembly of in vitro formed lamin head‐to‐tail polymers by CDC2 kinase. , 1991, The EMBO journal.

[8]  M. Peter,et al.  Expression of chicken lamin B2 in Escherichia coli: characterization of its structure, assembly, and molecular interactions , 1991, The Journal of cell biology.

[9]  C. Smythe,et al.  Assembly/disassembly of the nuclear envelope membrane: Cell cycle-dependent binding of nuclear membrane vesicles to chromatin in vitro , 1991, Cell.

[10]  D. Beach,et al.  A role for the p34cdc2 kinase and phosphatases in the regulation of phosphorylation and disassembly of lamin B2 during the cell cycle. , 1991, The EMBO journal.

[11]  D. Beach,et al.  A complex containing p34cdc2 and cyclin B phosphorylates the nuclear lamin and disassembles nuclei of clam oocytes in vitro , 1991, The Journal of cell biology.

[12]  B. A. Hocevar,et al.  Selective translocation of beta II-protein kinase C to the nucleus of human promyelocytic (HL60) leukemia cells. , 1991, The Journal of biological chemistry.

[13]  L. Lin,et al.  Immunoaffinity purification and functional characterization of interphase and meiotic Drosophila nuclear lamin isoforms. , 1990, Journal of Biological Chemistry.

[14]  F. McKeon,et al.  Mutations of phosphorylation sites in lamin A that prevent nuclear lamina disassembly in mitosis , 1990, Cell.

[15]  M. Kirschner,et al.  Identification of cell cycle-regulated phosphorylation sites on nuclear lamin C , 1990, Cell.

[16]  J. Labbé,et al.  In vitro disassembly of the nuclear lamina and M phase-specific phosphorylation of lamins by cdc2 kinase , 1990, Cell.

[17]  Michael Ashburner,et al.  Drosophila: A laboratory handbook , 1990 .

[18]  E. Nigg,et al.  A somatic cell-derived system for studying both early and late mitotic events in vitro. , 1989, Journal of cell science.

[19]  J. M. Lee,et al.  Heat shock-induced appearance of RNA polymerase II in karyoskeletal protein-enriched (nuclear "matrix") fractions correlates with transcriptional shutdown in Drosophila melanogaster. , 1989, The Journal of biological chemistry.

[20]  P. Fisher,et al.  Interconversion of Drosophila nuclear lamin isoforms during oogenesis, early embryogenesis, and upon entry of cultured cells into mitosis , 1989, The Journal of cell biology.

[21]  L. Rebhun,et al.  Disassembly of the nuclear envelope of spisula oocytes in a cell-free system. , 1989, Developmental biology.

[22]  D F Hochstrasser,et al.  Methods for increasing the resolution of two-dimensional protein electrophoresis. , 1988, Analytical biochemistry.

[23]  P. Fisher,et al.  Affinity purification of antibodies using antigens immobilized on solid supports. , 1988, Biochemical Society transactions.

[24]  J. Sedat,et al.  Drosophila nuclear lamin precursor Dm0 is translated from either of two developmentally regulated mRNA species apparently encoded by a single gene [published erratum appears in J Cell Biol 1988 Jun;106(6):2225] , 1988, Journal of Cell Biology.

[25]  David E. Smith,et al.  Heat shock-induced changes in the structural stability of proteinaceous karyoskeletal elements in vitro and morphological effects in situ , 1987, The Journal of cell biology.

[26]  Y. Gruenbaum,et al.  Biosynthesis and interconversion of Drosophila nuclear lamin isoforms during normal growth and in response to heat shock , 1987, The Journal of cell biology.

[27]  J. Newport,et al.  Disassembly of the nucleus in mitotic extracts: Membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes , 1987, Cell.

[28]  J. Newport.  Nuclear reconstitution in vitro: Stages of assembly around protein-free DNA , 1987, Cell.

[29]  L. Gerace,et al.  A fractionated cell-free system for analysis of prophase nuclear disassembly , 1986, The Journal of cell biology.

[30]  J. Maller,et al.  Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts , 1985, The Journal of cell biology.

[31]  M. Kirschner,et al.  Induction of early mitotic events in a cell-free system , 1985, Cell.

[32]  L. Gerace,et al.  Phosphorylation of the nuclear lamins during interphase and mitosis. , 1985, The Journal of biological chemistry.

[33]  P. Fisher,et al.  Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots , 1984, The Journal of cell biology.

[34]  M. Lohka,et al.  Effects of Ca2+ ions on the formation of metaphase chromosomes and sperm pronuclei in cell-free preparations from unactivated Rana pipiens eggs. , 1984, Developmental biology.

[35]  E. Gotschlich,et al.  A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. , 1984, Analytical biochemistry.

[36]  G. Blobel,et al.  Isolation and characterization of a proteinaceous subnuclear fraction composed of nuclear matrix, peripheral lamina, and nuclear pore complexes from embryos of Drosophila melanogaster , 1982, The Journal of cell biology.

[37]  G. Blobel,et al.  The nuclear envelope lamina is reversibly depolymerized during mitosis , 1980, Cell.

[38]  Y. Masui,et al.  Ca and Mg control of cytostatic factors from Rana pipiens oocytes which cause metaphase and cleavage arrest. , 1977, Developmental biology.

[39]  M. Crippa,et al.  Mass fractionation of Drosophila egg chambers. , 1977, Developmental biology.

[40]  C. Allis,et al.  Mass isolation of pole cells from Drosophila melanogaster. , 1977, Developmental biology.

[41]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.