Journal of Cell Science, Vol 106, Issue 1 275-285, Copyright © 1993 by Company of Biologists

JOURNAL ARTICLES

Nuclei that lack a lamina accumulate karyophilic proteins and assemble a nuclear matrix

H Jenkins, T Holman, C Lyon, B Lane, R Stick and C Hutchison
Department of Biological Sciences, The University, Dundee, UK.

Xenopus egg extracts, which support nuclear assembly and DNA replication in vitro, were physically depleted of lamin B3 using monoclonal antibodies linked to magnetic beads. Depleted extracts were still able to support nuclear envelope assembly around demembranated sperm heads but the resulting pronuclei lacked a lamina and were unable to initiate semiconservative DNA replication or to assemble replicases, confirming previous data. Immunoblotting analysis of isolated nuclei and nuclear matrix fractions indicated that lamin-depleted nuclei still accumulated nucleoporins and PCNA. Furthermore, the rate of PCNA uptake was identical in lamin-depleted and control nuclei. However, neither the nucleoporins nor the PCNA was associated with nuclear matrix fractions. The major protein components of sperm pronuclear matrix fractions were characterized by two-dimensional gel electrophoresis. Of these proteins only three out of 22 species, other than the lamins, were significantly reduced in lamin-depleted nuclei, indicating that these nuclei do assemble a nuclear matrix.

This article has been cited by other articles:

				D. K. Shumaker, L. Solimando, K. Sengupta, T. Shimi, S. A. Adam, A. Grunwald, S. V. Strelkov, U. Aebi, M. C. Cardoso, and R. D. Goldman The highly conserved nuclear lamin Ig-fold binds to PCNA: its role in DNA replication J. Cell Biol., April 21, 2008; 181(2): 269 - 280. [Abstract] [Full Text] [PDF]

				T. Dechat, K. Pfleghaar, K. Sengupta, T. Shimi, D. K. Shumaker, L. Solimando, and R. D. Goldman Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin Genes & Dev., April 1, 2008; 22(7): 832 - 853. [Abstract] [Full Text] [PDF]

				S. A. Adam, K. Sengupta, and R. D. Goldman Regulation of Nuclear Lamin Polymerization by Importin {alpha} J. Biol. Chem., March 28, 2008; 283(13): 8462 - 8468. [Abstract] [Full Text] [PDF]

				J. L. V. Broers, F. C. S. Ramaekers, G. Bonne, R. B. Yaou, and C. J. Hutchison Nuclear lamins: laminopathies and their role in premature ageing. Physiol Rev, July 1, 2006; 86(3): 967 - 1008. [Abstract] [Full Text] [PDF]

				S. W. Krauss, C. Chen, S. Penman, and R. Heald Nuclear actin and protein 4.1: Essential interactions during nuclear assembly in vitro PNAS, September 16, 2003; 100(19): 10752 - 10757. [Abstract] [Full Text] [PDF]

				E. Markiewicz, T. Dechat, R. Foisner, Roy. A Quinlan, and C. J. Hutchison Lamin A/C Binding Protein LAP2alpha Is Required for Nuclear Anchorage of Retinoblastoma Protein Mol. Biol. Cell, December 1, 2002; 13(12): 4401 - 4413. [Abstract] [Full Text] [PDF]

				O. A. Vaughan, M. Alvarez-Reyes, J. M. Bridger, J. L. V. Broers, F. C. S. Ramaekers, M. Wehnert, G. E. Morris, W. G. F. Whitfield, and C. J. Hutchison Both emerin and lamin C depend on lamin A for localization at the nuclear envelope J. Cell Sci., March 9, 2002; 114(14): 2577 - 2590. [Abstract] [Full Text] [PDF]

				R. D. Goldman, Y. Gruenbaum, R. D. Moir, D. K. Shumaker, and T. P. Spann Nuclear lamins: building blocks of nuclear architecture Genes & Dev., March 1, 2002; 16(5): 533 - 547. [Full Text] [PDF]

				E. A. L. Fairley, A. Riddell, J. A. Ellis, and J. Kendrick-Jones The cell cycle dependent mislocalisation of emerin may contribute to the Emery-Dreifuss muscular dystrophy phenotype J. Cell Sci., January 15, 2002; 115(2): 341 - 354. [Abstract] [Full Text] [PDF]

				R. I. Lopez-Soler, R. D. Moir, T. P. Spann, R. Stick, and R. D. Goldman A role for nuclear lamins in nuclear envelope assembly J. Cell Biol., July 9, 2001; 154(1): 61 - 70. [Abstract] [Full Text] [PDF]

				M. Izumi, O. A. Vaughan, C. J. Hutchison, and D. M. Gilbert Head and/or CaaX Domain Deletions of Lamin Proteins Disrupt Preformed Lamin A and C But Not Lamin B Structure in Mammalian Cells Mol. Biol. Cell, December 1, 2000; 11(12): 4323 - 4337. [Abstract] [Full Text]

				R. D. Moir, T. P. Spann, H. Herrmann, and R. D. Goldman Disruption of Nuclear Lamin Organization Blocks the Elongation Phase of DNA Replication J. Cell Biol., June 12, 2000; 149(6): 1179 - 1192. [Abstract] [Full Text] [PDF]

				J. Ortega and M. DePamphilis Nucleoskeleton and initiation of DNA replication in metazoan cells J. Cell Sci., June 14, 1999; 111(24): 3663 - 3673. [Abstract] [PDF]

				T. M. Gant, C. A. Harris, and K. L. Wilson Roles of LAP2 Proteins in Nuclear Assembly and DNA Replication: Truncated LAP2beta Proteins Alter Lamina Assembly, Envelope Formation, Nuclear Size, and DNA Replication Efficiency in Xenopus laevis Extracts J. Cell Biol., March 22, 1999; 144(6): 1083 - 1096. [Abstract] [Full Text] [PDF]

				S. Drummond, P. Ferrigno, C. Lyon, J. Murphy, M. Goldberg, T. Allen, C. Smythe, and C. J. Hutchison Temporal Differences in the Appearance of NEP-B78 and an LBR-like Protein during Xenopus Nuclear Envelope Reassembly Reflect the Ordered Recruitment of Functionally Discrete Vesicle Types J. Cell Biol., January 25, 1999; 144(2): 225 - 240. [Abstract] [Full Text] [PDF]

				P Collas Nuclear envelope disassembly in mitotic extract requires functional nuclear pores and a nuclear lamina J. Cell Sci., January 5, 1998; 111(9): 1293 - 1303. [Abstract] [PDF]

				B. Lenz-Bohme, J. Wismar, S. Fuchs, R. Reifegerste, E. Buchner, H. Betz, and B. Schmitt Insertional Mutation of the Drosophila Nuclear Lamin Dm0 Gene Results in Defective Nuclear Envelopes, Clustering of Nuclear Pore Complexes, and Accumulation of Annulate Lamellae J. Cell Biol., June 2, 1997; 137(5): 1001 - 1016. [Abstract] [Full Text] [PDF]

				T. P. Spann, R. D. Moir, A. E. Goldman, R. Stick, and R. D. Goldman Disruption of Nuclear Lamin Organization Alters the Distribution of Replication Factors and Inhibits DNA Synthesis J. Cell Biol., March 24, 1997; 136(6): 1201 - 1212. [Abstract] [Full Text] [PDF]

				Z. Lu, D. Sittman, D. Brown, R Munshi, and G. Leno Histone H1 modulates DNA replication through multiple pathways in Xenopus egg extract J. Cell Sci., January 11, 1997; 110(21): 2745 - 2758. [Abstract] [PDF]

				D. Ellis, H Jenkins, W. Whitfield, and C. Hutchison GST-lamin fusion proteins act as dominant negative mutants in Xenopus egg extract and reveal the function of the lamina in DNA replication J. Cell Sci., January 10, 1997; 110(20): 2507 - 2518. [Abstract] [PDF]

				C Zhang, H Jenkins, M. Goldberg, T. Allen, and C. Hutchison Nuclear lamina and nuclear matrix organization in sperm pronuclei assembled in Xenopus egg extract J. Cell Sci., January 9, 1996; 109(9): 2275 - 2286. [Abstract] [PDF]

				M Goldberg, H Jenkins, T Allen, W. Whitfield, and C. Hutchison Xenopus lamin B3 has a direct role in the assembly of a replication competent nucleus: evidence from cell-free egg extracts J. Cell Sci., January 11, 1995; 108(11): 3451 - 3461. [Abstract] [PDF]

				S Chevalier, J. Tassan, R Cox, M Philippe, and C Ford Both cdc2 and cdk2 promote S phase initiation in Xenopus egg extracts J. Cell Sci., January 5, 1995; 108(5): 1831 - 1841. [Abstract] [PDF]

				P Hozak, A. Sasseville, Y Raymond, and P. Cook Lamin proteins form an internal nucleoskeleton as well as a peripheral lamina in human cells J. Cell Sci., January 2, 1995; 108(2): 635 - 644. [Abstract] [PDF]

				J Murphy, C. Crompton, S Hainey, G. Codd, and C. Hutchison The role of protein phosphorylation in the assembly of a replication competent nucleus: investigations in Xenopus egg extracts using the cyanobacterial toxin microcystin-LR J. Cell Sci., January 1, 1995; 108(1): 235 - 244. [Abstract] [PDF]

				C. Hutchison, J. Bridger, L. Cox, and I. Kill Weaving a pattern from disparate threads: lamin function in nuclear assembly and DNA replication J. Cell Sci., January 12, 1994; 107(12): 3259 - 3269. [Abstract] [PDF]