Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1995 Jul 17;14(14):3311–3324. doi: 10.1002/j.1460-2075.1995.tb07338.x

Vimentin-associated mitotic vesicles interact with chromosomes in a lamin B- and phosphorylation-dependent manner.

C Maison 1, A Pyrpasopoulou 1, S D Georgatos 1
PMCID: PMC394399  PMID: 7628433

Abstract

We have assessed the involvement of the nuclear lamins in nuclear envelope reassembly. Analysis of perforated mitotic cells shows that A-type lamins are partly cytosolic and partly chromosome-bound, whereas B-type lamins are associated with vesicular structures throughout cell division. Lamin B-containing vesicles appear to dock on vimentin intermediate filaments during prometaphase, but dissociate from the cytoskeleton and assemble around chromatin at later phases of mitosis. Mitotic vesicles isolated from prometaphase cells en bloc with vimentin filaments can specifically capture chromosomes. Efficient chromosome capturing requires cytosolic factors and a dephosphorylating environment. Urea-stripping of the vesicles abolishes binding to chromosomes. However, reconstitution of the stripped membranes with purified B-type lamins restores their ability to bind to chromosomes in a cytosol- and dephosphorylation-dependent fashion. Vesicles reconstituted with B-type lamins form membraneous 'crescents' on the surfaces of chromosomes, but, unlike native vesicles, do not fuse into large sheets. From these observations we conclude that the initial targeting of mitotic vesicles to chromosomes is dependent on B-type lamins and on factors present in the mitotic cytoplasm. Apparently, further recruitment of membranes and fusion of chromosome-bound vesicles onto chromatin involves non-lamin peripheral membrane proteins.

Full text

PDF
3311

Images in this article

3312Image
on p.3312
3313Image
on p.3313
3314Image
on p.3314
3315Image
on p.3315
3317Image
on p.3317
3317Image
on p.3317
3318Image
on p.3318
3320Image
on p.3320
3321Image
on p.3321

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bailly E., Dorée M., Nurse P., Bornens M. p34cdc2 is located in both nucleus and cytoplasm; part is centrosomally associated at G2/M and enters vesicles at anaphase. EMBO J. 1989 Dec 20;8(13):3985–3995. doi: 10.1002/j.1460-2075.1989.tb08581.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blobel G., Dobberstein B. Transfer of proteins across membranes. II. Reconstitution of functional rough microsomes from heterologous components. J Cell Biol. 1975 Dec;67(3):852–862. doi: 10.1083/jcb.67.3.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boman A. L., Delannoy M. R., Wilson K. L. GTP hydrolysis is required for vesicle fusion during nuclear envelope assembly in vitro. J Cell Biol. 1992 Jan;116(2):281–294. doi: 10.1083/jcb.116.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boman A. L., Taylor T. C., Melançon P., Wilson K. L. A role for ADP-ribosylation factor in nuclear vesicle dynamics. Nature. 1992 Aug 6;358(6386):512–514. doi: 10.1038/358512a0. [DOI] [PubMed] [Google Scholar]
  5. Burke B., Gerace L. A cell free system to study reassembly of the nuclear envelope at the end of mitosis. Cell. 1986 Feb 28;44(4):639–652. doi: 10.1016/0092-8674(86)90273-4. [DOI] [PubMed] [Google Scholar]
  6. Chaudhary N., Courvalin J. C. Stepwise reassembly of the nuclear envelope at the end of mitosis. J Cell Biol. 1993 Jul;122(2):295–306. doi: 10.1083/jcb.122.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Colucci-Guyon E., Portier M. M., Dunia I., Paulin D., Pournin S., Babinet C. Mice lacking vimentin develop and reproduce without an obvious phenotype. Cell. 1994 Nov 18;79(4):679–694. doi: 10.1016/0092-8674(94)90553-3. [DOI] [PubMed] [Google Scholar]
  8. Duncan J. L., Schlegel R. Effect of streptolysin O on erythrocyte membranes, liposomes, and lipid dispersions. A protein-cholesterol interaction. J Cell Biol. 1975 Oct;67(1):160–174. doi: 10.1083/jcb.67.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dwyer N., Blobel G. A modified procedure for the isolation of a pore complex-lamina fraction from rat liver nuclei. J Cell Biol. 1976 Sep;70(3):581–591. doi: 10.1083/jcb.70.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Foisner R., Gerace L. Integral membrane proteins of the nuclear envelope interact with lamins and chromosomes, and binding is modulated by mitotic phosphorylation. Cell. 1993 Jul 2;73(7):1267–1279. doi: 10.1016/0092-8674(93)90355-t. [DOI] [PubMed] [Google Scholar]
  11. Georgatos S. D., Blobel G. Lamin B constitutes an intermediate filament attachment site at the nuclear envelope. J Cell Biol. 1987 Jul;105(1):117–125. doi: 10.1083/jcb.105.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Georgatos S. D., Meier J., Simos G. Lamins and lamin-associated proteins. Curr Opin Cell Biol. 1994 Jun;6(3):347–353. doi: 10.1016/0955-0674(94)90025-6. [DOI] [PubMed] [Google Scholar]
  13. Georgatos S. D., Stournaras C., Blobel G. Heterotypic and homotypic associations between the nuclear lamins: site-specificity and control by phosphorylation. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4325–4329. doi: 10.1073/pnas.85.12.4325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Georgatos S. D., Weber K., Geisler N., Blobel G. Binding of two desmin derivatives to the plasma membrane and the nuclear envelope of avian erythrocytes: evidence for a conserved site-specificity in intermediate filament-membrane interactions. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6780–6784. doi: 10.1073/pnas.84.19.6780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gerace L., Blobel G. The nuclear envelope lamina is reversibly depolymerized during mitosis. Cell. 1980 Jan;19(1):277–287. doi: 10.1016/0092-8674(80)90409-2. [DOI] [PubMed] [Google Scholar]
  16. Glass J. R., Gerace L. Lamins A and C bind and assemble at the surface of mitotic chromosomes. J Cell Biol. 1990 Sep;111(3):1047–1057. doi: 10.1083/jcb.111.3.1047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gravotta D., Adesnik M., Sabatini D. D. Transport of influenza HA from the trans-Golgi network to the apical surface of MDCK cells permeabilized in their basolateral plasma membranes: energy dependence and involvement of GTP-binding proteins. J Cell Biol. 1990 Dec;111(6 Pt 2):2893–2908. doi: 10.1083/jcb.111.6.2893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Heald R., McKeon F. Mutations of phosphorylation sites in lamin A that prevent nuclear lamina disassembly in mitosis. Cell. 1990 May 18;61(4):579–589. doi: 10.1016/0092-8674(90)90470-y. [DOI] [PubMed] [Google Scholar]
  19. Höger T. H., Krohne G., Kleinschmidt J. A. Interaction of Xenopus lamins A and LII with chromatin in vitro mediated by a sequence element in the carboxyterminal domain. Exp Cell Res. 1991 Dec;197(2):280–289. doi: 10.1016/0014-4827(91)90434-v. [DOI] [PubMed] [Google Scholar]
  20. Kouklis P. D., Merdes A., Papamarcaki T., Georgatos S. D. Transient arrest of 3T3 cells in mitosis and inhibition of nuclear lamin reassembly around chromatin induced by anti-vimentin antibodies. Eur J Cell Biol. 1993 Dec;62(2):224–236. [PubMed] [Google Scholar]
  21. Krohne G., Wolin S. L., McKeon F. D., Franke W. W., Kirschner M. W. Nuclear lamin LI of Xenopus laevis: cDNA cloning, amino acid sequence and binding specificity of a member of the lamin B subfamily. EMBO J. 1987 Dec 1;6(12):3801–3808. doi: 10.1002/j.1460-2075.1987.tb02716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lourim D., Krohne G. Lamin-dependent nuclear envelope reassembly following mitosis: an argument. Trends Cell Biol. 1994 Sep;4(9):314–318. doi: 10.1016/0962-8924(94)90228-3. [DOI] [PubMed] [Google Scholar]
  24. Ludérus M. E., de Graaf A., Mattia E., den Blaauwen J. L., Grande M. A., de Jong L., van Driel R. Binding of matrix attachment regions to lamin B1. Cell. 1992 Sep 18;70(6):949–959. doi: 10.1016/0092-8674(92)90245-8. [DOI] [PubMed] [Google Scholar]
  25. Maison C., Horstmann H., Georgatos S. D. Regulated docking of nuclear membrane vesicles to vimentin filaments during mitosis. J Cell Biol. 1993 Dec;123(6 Pt 1):1491–1505. doi: 10.1083/jcb.123.6.1491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Meier J., Georgatos S. D. Type B lamins remain associated with the integral nuclear envelope protein p58 during mitosis: implications for nuclear reassembly. EMBO J. 1994 Apr 15;13(8):1888–1898. doi: 10.1002/j.1460-2075.1994.tb06458.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Newport J., Dunphy W. Characterization of the membrane binding and fusion events during nuclear envelope assembly using purified components. J Cell Biol. 1992 Jan;116(2):295–306. doi: 10.1083/jcb.116.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Papamarcaki T., Kouklis P. D., Kreis T. E., Georgatos S. D. The "lamin B-fold". Anti-idiotypic antibodies reveal a structural complementarity between nuclear lamin B and cytoplasmic intermediate filament epitopes. J Biol Chem. 1991 Nov 5;266(31):21247–21251. [PubMed] [Google Scholar]
  29. Peter M., Heitlinger E., Häner M., Aebi U., Nigg E. A. Disassembly of in vitro formed lamin head-to-tail polymers by CDC2 kinase. EMBO J. 1991 Jun;10(6):1535–1544. doi: 10.1002/j.1460-2075.1991.tb07673.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Peter M., Nakagawa J., Dorée M., Labbé J. C., Nigg E. A. In vitro disassembly of the nuclear lamina and M phase-specific phosphorylation of lamins by cdc2 kinase. Cell. 1990 May 18;61(4):591–602. doi: 10.1016/0092-8674(90)90471-p. [DOI] [PubMed] [Google Scholar]
  31. Pfaller R., Smythe C., Newport J. W. Assembly/disassembly of the nuclear envelope membrane: cell cycle-dependent binding of nuclear membrane vesicles to chromatin in vitro. Cell. 1991 Apr 19;65(2):209–217. doi: 10.1016/0092-8674(91)90155-r. [DOI] [PubMed] [Google Scholar]
  32. Pimplikar S. W., Ikonen E., Simons K. Basolateral protein transport in streptolysin O-permeabilized MDCK cells. J Cell Biol. 1994 Jun;125(5):1025–1035. doi: 10.1083/jcb.125.5.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sarria A. J., Lieber J. G., Nordeen S. K., Evans R. M. The presence or absence of a vimentin-type intermediate filament network affects the shape of the nucleus in human SW-13 cells. J Cell Sci. 1994 Jun;107(Pt 6):1593–1607. doi: 10.1242/jcs.107.6.1593. [DOI] [PubMed] [Google Scholar]
  34. Simos G., Georgatos S. D. The inner nuclear membrane protein p58 associates in vivo with a p58 kinase and the nuclear lamins. EMBO J. 1992 Nov;11(11):4027–4036. doi: 10.1002/j.1460-2075.1992.tb05496.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stewart C., Burke B. Teratocarcinoma stem cells and early mouse embryos contain only a single major lamin polypeptide closely resembling lamin B. Cell. 1987 Nov 6;51(3):383–392. doi: 10.1016/0092-8674(87)90634-9. [DOI] [PubMed] [Google Scholar]
  36. Sullivan K. M., Busa W. B., Wilson K. L. Calcium mobilization is required for nuclear vesicle fusion in vitro: implications for membrane traffic and IP3 receptor function. Cell. 1993 Jul 2;73(7):1411–1422. doi: 10.1016/0092-8674(93)90366-x. [DOI] [PubMed] [Google Scholar]
  37. Suprynowicz F. A., Gerace L. A fractionated cell-free system for analysis of prophase nuclear disassembly. J Cell Biol. 1986 Dec;103(6 Pt 1):2073–2081. doi: 10.1083/jcb.103.6.2073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Suprynowicz F. A. Inactivation of cdc2 kinase during mitosis requires regulated and constitutive proteins in a cell-free system. J Cell Sci. 1993 Mar;104(Pt 3):873–881. doi: 10.1242/jcs.104.3.873. [DOI] [PubMed] [Google Scholar]
  39. Vigers G. P., Lohka M. J. A distinct vesicle population targets membranes and pore complexes to the nuclear envelope in Xenopus eggs. J Cell Biol. 1991 Feb;112(4):545–556. doi: 10.1083/jcb.112.4.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ward G. E., Kirschner M. W. Identification of cell cycle-regulated phosphorylation sites on nuclear lamin C. Cell. 1990 May 18;61(4):561–577. doi: 10.1016/0092-8674(90)90469-u. [DOI] [PubMed] [Google Scholar]
  41. Wiese C., Wilson K. L. Nuclear membrane dynamics. Curr Opin Cell Biol. 1993 Jun;5(3):387–394. doi: 10.1016/0955-0674(93)90002-8. [DOI] [PubMed] [Google Scholar]
  42. Wilson K. L., Newport J. A trypsin-sensitive receptor on membrane vesicles is required for nuclear envelope formation in vitro. J Cell Biol. 1988 Jul;107(1):57–68. doi: 10.1083/jcb.107.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Worman H. J., Evans C. D., Blobel G. The lamin B receptor of the nuclear envelope inner membrane: a polytopic protein with eight potential transmembrane domains. J Cell Biol. 1990 Oct;111(4):1535–1542. doi: 10.1083/jcb.111.4.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Worman H. J., Yuan J., Blobel G., Georgatos S. D. A lamin B receptor in the nuclear envelope. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8531–8534. doi: 10.1073/pnas.85.22.8531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yuan J., Simos G., Blobel G., Georgatos S. D. Binding of lamin A to polynucleosomes. J Biol Chem. 1991 May 15;266(14):9211–9215. [PubMed] [Google Scholar]
  46. Zieve G. W., Turnbull D., Mullins J. M., McIntosh J. R. Production of large numbers of mitotic mammalian cells by use of the reversible microtubule inhibitor nocodazole. Nocodazole accumulated mitotic cells. Exp Cell Res. 1980 Apr;126(2):397–405. doi: 10.1016/0014-4827(80)90279-7. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES