Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 499K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Moir, R. D. Articles by Goldman, R. D. Search for Related Content PubMed PubMed Citation Articles by Moir, R. D. Articles by Goldman, R. D. Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/2000//1179 $5.00
The Journal of Cell Biology, Volume 149, Number 6, , 2000 1179-1192

Disruption of Nuclear Lamin Organization Blocks the Elongation Phase of DNA Replication

^a Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611
^b Division of Cell Biology, German Cancer Research Center, D-69120 Heidelberg, Germany
Department of Cell and Molecular Biology, Northwestern University Medical School, 303 E. Chicago Ave., Ward 11-145, Chicago, IL 60611.(312) 503-0954(312) 503-4215

r-goldman{at}nwu.edu

The role of nuclear lamins in DNA replication is unclear. To address this, nuclei were assembled in Xenopus extracts containing AraC, a reversible inhibitor that blocks near the onset of the elongation phase of replication. Dominant-negative lamin mutants lacking their NH₂-terminal domains were added to assembled nuclei to disrupt lamin organization. This prevented the resumption of DNA replication after the release of the AraC block. This inhibition of replication was not due to gross disruption of nuclear envelope structure and function. The organization of initiation factors was not altered by lamin disruption, and nuclei resumed replication when transferred to extracts treated with CIP, an inhibitor of the cyclin-dependent kinase (cdk) 2–dependent step of initiation. This suggests that alteration of lamin organization does not affect the initiation phase of DNA replication. Instead, we find that disruption of lamin organization inhibited chain elongation in a dose-dependent fashion. Furthermore, the established organization of two elongation factors, proliferating cell nuclear antigen, and replication factor complex, was disrupted by NLA. These findings demonstrate that lamin organization must be maintained in nuclei for the elongation phase of DNA replication to proceed.

Key Words: DNA synthesis • lamina • intermediate filaments • nuclear organization • lamin mutant

© 2000 The Rockefeller University Press

Abbreviations used in this paper: cdk, cyclin-dependent kinase; GST, glutathione S-transferase; LA, human nuclear lamin A; LB3, Xenopus nuclear lamin B3; NLS, nuclear localization signal; NWB, nuclear wash buffer; PCNA, proliferating cell nuclear antigen; RFC, replication factor complex; XMCM, Xenopus minichromosome maintenance; factor XORC2, Xenopus origin replication complex factor 2.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Facebook

Reddit

Technorati

Twitter

This article has been cited by other articles:

• Arnault, E., Doussau, M., Pesty, A., Lefevre, B., Courtot, A.-M. (2010). Review: Lamin A/C, Caspase-6, and Chromatin Configuration During Meiosis Resumption in the Mouse Oocyte. Reproductive Sciences 17: 102-115 [Abstract]  
• Graham, J. D., Mote, P. A., Salagame, U., van Dijk, J. H., Balleine, R. L., Huschtscha, L. I., Reddel, R. R., Clarke, C. L. (2009). DNA Replication Licensing and Progenitor Numbers Are Increased by Progesterone in Normal Human Breast. Endocrinology 150: 3318-3326 [Abstract] [Full Text]  
• Krasinska, L., Fisher, D. (2009). Replication initiation complex formation in the absence of nuclear function in Xenopus. Nucleic Acids Res 37: 2238-2248 [Abstract] [Full Text]  
• Shumaker, D. K., Solimando, L., Sengupta, K., Shimi, T., Adam, S. A., Grunwald, A., Strelkov, S. V., Aebi, U., Cardoso, M. C., Goldman, R. D. (2008). The highly conserved nuclear lamin Ig-fold binds to PCNA: its role in DNA replication. JCB 181: 269-280 [Abstract] [Full Text]  
• Dechat, T., Pfleghaar, K., Sengupta, K., Shimi, T., Shumaker, D. K., Solimando, L., Goldman, R. D. (2008). Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin. Genes Dev. 22: 832-853 [Abstract] [Full Text]  
• Liu, Y., Wang, Y., Rusinol, A. E., Sinensky, M. S., Liu, J., Shell, S. M., Zou, Y. (2008). Involvement of xeroderma pigmentosum group A (XPA) in progeria arising from defective maturation of prelamin A. FASEB J. 22: 603-611 [Abstract] [Full Text]  
• Muchir, A., Pavlidis, P., Bonne, G., Hayashi, Y. K., Worman, H. J. (2007). Activation of MAPK in hearts of EMD null mice: similarities between mouse models of X-linked and autosomal dominant Emery Dreifuss muscular dystrophy. Hum Mol Genet 16: 1884-1895 [Abstract] [Full Text]  
• Naetar, N., Hutter, S., Dorner, D., Dechat, T., Korbei, B., Gotzmann, J., Beug, H., Foisner, R. (2007). LAP2{alpha}-binding protein LINT-25 is a novel chromatin-associated protein involved in cell cycle exit. J. Cell Sci. 120: 737-747 [Abstract] [Full Text]  
• Manju, K., Muralikrishna, B., Parnaik, V. K (2006). Expression of disease-causing lamin A mutants impairs the formation of DNA repair foci. J. Cell Sci. 119: 2704-2714 [Abstract] [Full Text]  
• Dorner, D., Vlcek, S., Foeger, N., Gajewski, A., Makolm, C., Gotzmann, J., Hutchison, C. J., Foisner, R. (2006). Lamina-associated polypeptide 2{alpha} regulates cell cycle progression and differentiation via the retinoblastoma-E2F pathway.. JCB 173: 83-93 [Abstract] [Full Text]  
• Tsai, M.-Y., Wang, S., Heidinger, J. M., Shumaker, D. K., Adam, S. A., Goldman, R. D., Zheng, Y. (2006). A Mitotic Lamin B Matrix Induced by RanGTP Required for Spindle Assembly. Science 311: 1887-1893 [Abstract] [Full Text]  
• Boguslavsky, R. L., Stewart, C. L., Worman, H. J. (2006). Nuclear lamin A inhibits adipocyte differentiation: implications for Dunnigan-type familial partial lipodystrophy. Hum Mol Genet 15: 653-663 [Abstract] [Full Text]  
• Shumaker, D. K., Lopez-Soler, R. I., Adam, S. A., Herrmann, H., Moir, R. D., Spann, T. P., Goldman, R. D. (2005). Functions and dysfunctions of the nuclear lamin Ig-fold domain in nuclear assembly, growth, and Emery-Dreifuss muscular dystrophy. Proc. Natl. Acad. Sci. USA 102: 15494-15499 [Abstract] [Full Text]  
• Mariappan, I., Parnaik, V. K. (2005). Sequestration of pRb by Cyclin D3 Causes Intranuclear Reorganization of Lamin A/C during Muscle Cell Differentiation. Mol. Biol. Cell 16: 1948-1960 [Abstract] [Full Text]  
• Markiewicz, E., Ledran, M., Hutchison, C. J. (2005). Remodelling of the nuclear lamina and nucleoskeleton is required for skeletal muscle differentiation in vitro. J. Cell Sci. 118: 409-420 [Abstract] [Full Text]  
• Broers, J. L.V., Peeters, E. A.G., Kuijpers, H. J.H., Endert, J., Bouten, C. V.C., Oomens, C. W.J., Baaijens, F. P.T., Ramaekers, F. C.S. (2004). Decreased mechanical stiffness in LMNA-/- cells is caused by defective nucleo-cytoskeletal integrity: implications for the development of laminopathies. Hum Mol Genet 13: 2567-2580 [Abstract] [Full Text]  
• Panorchan, P., Schafer, B. W., Wirtz, D., Tseng, Y. (2004). Nuclear Envelope Breakdown Requires Overcoming the Mechanical Integrity of the Nuclear Lamina. J. Biol. Chem. 279: 43462-43467 [Abstract] [Full Text]  
• Muchir, A., Worman, H. J. (2004). The Nuclear Envelope and Human Disease. Physiology 19: 309-314 [Abstract] [Full Text]  
• Veluthakal, R., Amin, R., Kowluru, A. (2004). Interleukin-1{beta} induces posttranslational carboxymethylation and alterations in subnuclear distribution of lamin B in insulin-secreting RINm5F cells. Am. J. Physiol. Cell Physiol. 287: C1152-C1162 [Abstract] [Full Text]  
• Gajewski, A., Csaszar, E., Foisner, R. (2004). A Phosphorylation Cluster in the Chromatin-binding Region Regulates Chromosome Association of LAP2{alpha}. J. Biol. Chem. 279: 35813-35821 [Abstract] [Full Text]  
• Hoyt, C. C., Bouchard, R. J., Tyler, K. L. (2004). Novel Nuclear Herniations Induced by Nuclear Localization of a Viral Protein. J. Virol. 78: 6360-6369 [Abstract] [Full Text]  
• Moreira, P. N., Robl, J. M., Collas, P. (2003). Architectural defects in pronuclei of mouse nuclear transplant embryos. J. Cell Sci. 116: 3713-3720 [Abstract] [Full Text]  
• Fischer, A. H., Taysavang, P., Jhiang, S. M. (2003). Nuclear Envelope Irregularity Is Induced by RET/PTC During Interphase. Am. J. Pathol. 163: 1091-1100 [Abstract] [Full Text]  
• Steen, R. L., Beullens, M., Landsverk, H. B., Bollen, M., Collas, P. (2003). AKAP149 is a novel PP1 specifier required to maintain nuclear envelope integrity in G1 phase. J. Cell Sci. 116: 2237-2246 [Abstract] [Full Text]  
• Martins, S., Eikvar, S., Furukawa, K., Collas, P. (2003). HA95 and LAP2{beta} mediate a novel chromatin-nuclear envelope interaction implicated in initiation of DNA replication. JCB 160: 177-188 [Abstract] [Full Text]  
• Kumaran, R. I., Muralikrishna, B., Parnaik, V. K. (2002). Lamin A/C speckles mediate spatial organization of splicing factor compartments and RNA polymerase II transcription. JCB 159: 783-793 [Abstract] [Full Text]  
• Elias, M. C. Q. B., Faria, M., Mortara, R. A., Motta, M. C. M., de Souza, W., Thiry, M., Schenkman, S. (2002). Chromosome Localization Changes in the Trypanosoma cruzi Nucleus. Eukaryot Cell 1: 944-953 [Abstract] [Full Text]  
• Markiewicz, E., Dechat, T., Foisner, R., Quinlan, Roy. A, Hutchison, C. J. (2002). Lamin A/C Binding Protein LAP2alpha Is Required for Nuclear Anchorage of Retinoblastoma Protein. Mol. Biol. Cell 13: 4401-4413 [Abstract] [Full Text]  
• Geles, K. G., Johnson, J. J., Jong, S., Adam, S. A. (2002). A Role for Caenorhabditis elegans Importin IMA-2 in Germ Line and Embryonic Mitosis. Mol. Biol. Cell 13: 3138-3147 [Abstract] [Full Text]  
• Vlcek, S., Korbei, B., Foisner, R. (2002). Distinct Functions of the Unique C Terminus of LAP2alpha in Cell Proliferation and Nuclear Assembly. J. Biol. Chem. 277: 18898-18907 [Abstract] [Full Text]  
• Vigouroux, C., Auclair, M., Dubosclard, E., Pouchelet, M., Capeau, J., Courvalin, J.-C., Buendia, B. (2002). Nuclear envelope disorganization in fibroblasts from lipodystrophic patients with heterozygous R482Q/W mutations in the lamin A/C gene. J. Cell Sci. 114: 4459-4468 [Abstract] [Full Text]  
• Muralikrishna, Bh., Dhawan, J., Rangaraj, N., Parnaik, V. K. (2002). Distinct changes in intranuclear lamin A/C organization during myoblast differentiation. J. Cell Sci. 114: 4001-4011 [Abstract] [Full Text]  
• Goldman, R. D., Gruenbaum, Y., Moir, R. D., Shumaker, D. K., Spann, T. P. (2002). Nuclear lamins: building blocks of nuclear architecture. Genes Dev. 16: 533-547 [Full Text]  
• Spann, T. P., Goldman, A. E., Wang, C., Huang, S., Goldman, R. D. (2002). Alteration of nuclear lamin organization inhibits RNA polymerase II-dependent transcription. JCB 156: 603-608 [Abstract] [Full Text]  
• Fairley, E. A. L., Riddell, A., Ellis, J. A., Kendrick-Jones, J. (2002). The cell cycle dependent mislocalisation of emerin may contribute to the Emery-Dreifuss muscular dystrophy phenotype. J. Cell Sci. 115: 341-354 [Abstract] [Full Text]  
• de Noronha, C. M. C., Sherman, M. P., Lin, H. W., Cavrois, M. V., Moir, R. D., Goldman, R. D., Greene, W. C. (2001). Dynamic Disruptions in Nuclear Envelope Architecture and Integrity Induced by HIV-1 Vpr. Science 294: 1105-1108 [Abstract] [Full Text]  
• Moir, R. D., Yoon, M., Khuon, S., Goldman, R. D. (2000). Nuclear Lamins a and B1: Different Pathways of Assembly during Nuclear Envelope Formation in Living Cells. JCB 151: 1155-1168 [Abstract] [Full Text]  
• Izumi, M., Vaughan, O. A., Hutchison, C. J., Gilbert, D. M. (2000). 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 11: 4323-4337 [Abstract] [Full Text]  
• Kennedy, B. K., Barbie, D. A., Classon, M., Dyson, N., Harlow, E. (2000). Nuclear organization of DNA replication in primary mammalian cells. Genes Dev. 14: 2855-2868 [Abstract] [Full Text]  
• Anglana, M., Debatisse, M. (2001). Dual Control of Replication Timing. STOCHASTIC ONSET BUT PROGRAMMED COMPLETION OF MAMMALIAN CHROMOSOME DUPLICATION. J. Biol. Chem. 276: 36639-36646 [Abstract] [Full Text]  
• Spann, T. P., Goldman, A. E., Wang, C., Huang, S., Goldman, R. D. (2002). Alteration of nuclear lamin organization inhibits RNA polymerase II-dependent transcription. JCB 156: 603-608 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents