Anchoring of heterochromatin to the nuclear envelope appears to be an important process ensuring the spatial organization of the chromatin structure and genome function in eukaryotic nuclei. Proteins of the inner nuclear membrane (INM) mediating these interactions are able to recognize lamina-associated heterochromatin domains (termed LAD) and simultaneously bind either lamin A/C or lamin B1. One of these proteins is the lamin B receptor (LBR) that binds lamin B1 and tethers heterochromatin to the INM in embryonic and undifferentiated cells. It is replaced by lamin A/C with specific lamin A/C binding proteins at the beginning of cell differentiation and in differentiated cells. Our functional experiments in cancer cell lines show that heterochromatin in cancer cells is tethered to the INM by LBR, which is downregulated together with lamin B1 at the onset of cell transition to senescence. The downregulation of these proteins in senescent cells leads to the detachment of centromeric repetitive sequences from INM, their relocation to the nucleoplasm, and distension. In cells, the expression of LBR and LB1 is highly coordinated as evidenced by the reduction of both proteins in LBR shRNA lines. The loss of the constitutive heterochromatin structure containing LADs results in changes in chromatin architecture and genome function and can be the reason for the permanent loss of cell proliferation in senescence.

Department of Cell Biology and Radiobiology Institute of Biophysics Czech Academy of Sciences Královopolská 135 Brno 61265 Czech Republic

Department of Molecular Epigenetics Institute of Biophysics Czech Academy of Sciences Královopolská 135 Brno 61265 Czech Republic

Zobrazit více v PubMed

Woodcock C.L., Ghosh R.P. Chromatin higher order structure and dynamics. Cold Spring Harb. Perspect. Biol. 2010 doi: 10.1101/cshperspect.a000596. PubMed DOI PMC

Guelen L., Pagie L., Brasset E., Meuleman W., Faza M.B., Talhout W., Eussen B.H., de Klein A., Wessels L., de Laat W., et al. Domain organization of human chromosomes revealed by mapping of nuclear lamina interactions in single human cells. Nature. 2008;453:948–951. doi: 10.1038/nature06947. PubMed DOI

Solovei I., Wang A.S., Thanisch K., Schmidt C.S., Krebs S., Zwerger M., Cohen T.V., Devys D., Foisner R., Peichl L., et al. LBR and lamin A/C sequentially tether peripheral heterochromatin and inversely regulate differentiation. Cell. 2013;152:584–598. doi: 10.1016/j.cell.2013.01.009. PubMed DOI

Andrés V., González J.M. Role of A-type lamins in signaling, transcription, and chromatin organization. J. Cell Biol. 2009;187:945–957. doi: 10.1083/jcb.200904124. PubMed DOI PMC

Wagner N., Krohne G. LEM-Domain proteins: New insights into lamin-interacting proteins. Int. Rev. Cytol. 2007;216:1–46. PubMed

Brachner A., Foisner R. Evolvement of LEM proteins as chromatin tethers at the nuclear periphery. Biochem. Soc. Trans. 2011;39:1735–1741. doi: 10.1042/BST20110724. PubMed DOI PMC

Makatsori D., Kourmouli N., Polioudaki H., Shultz L.D., McLean K., Theodoropoulos P.A., Singh P.B., Georgatos S.D. The inner nuclear membrane protein lamin B receptor forms distinct microdomains and links epigenetically marked chromatin to the nuclear envelope. J. Biol. Chem. 2004;279:25567–25573. doi: 10.1074/jbc.M313606200. PubMed DOI

Olins A.L., Rhodes G., Welch D.B., Zwerger M., Olins D.E. Lamin B receptor: Multi-tasking at the nuclear envelope. Nucleus. 2010;1:53–70. doi: 10.4161/nucl.1.1.10515. PubMed DOI PMC

Hirano Y., Hizume K., Kimura H., Takeyasu K., Haraguchi T., Hiraoka Y. Lamin B receptor recognizes specific modifications of histone H4 in heterochromatin formation. J. Biol. Chem. 2012;287:42654–42663. doi: 10.1074/jbc.M112.397950. PubMed DOI PMC

Clowney E.J., LeGros M.A., Mosley C.P., Clowney F.G., Markenskoff-Papadimitriou E.C., Myllys M., Barnea G., Larabell C.A., Lomvardas S. Nuclear aggregation of olfactory receptor genes governs their monogenic expression. Cell. 2012;151:724–737. doi: 10.1016/j.cell.2012.09.043. PubMed DOI PMC

Kim Y., Sharov A.A., McDole K., Cheng M., Hao A., Fan C.M., Giano M.N., Ko M.S., Zheng Y. Mouse B-type lamins are required for proper organogenesis but not by embryonic stem cells. Science. 2011;334:1706–1710. doi: 10.1126/science.1211222. PubMed DOI PMC

Yang S.H., Chang S.Y., Yin L., Tu Y., Hu Y., Yoshinaga Y., de Jong P.J., Fong L.G., Young S.G. An absence of both lamin B1 and lamin B2 in keratinocytes has no effect on cell proliferation or the development of skin and hair. Hum. Mol. Genet. 2011;20:3537–3544. doi: 10.1093/hmg/ddr266. PubMed DOI PMC

Broers J.L., Ramaekers F.C., Bonne G., Yaou R.B., Hutchison C.J. Nuclear lamins: Laminopathies and their role in premature ageing. Phisiol. Rev. 2006;86:967–1008. doi: 10.1152/physrev.00047.2005. PubMed DOI

Holmer L., Worman H.J. Inner nuclear membrane proteins: Functions and targeting. Cell. Mol. Life Sci. 2001;58:1741–1747. doi: 10.1007/PL00000813. PubMed DOI PMC

Ikegami K., Egelhofer T.A., Strome S., Lieb J.D. Caenorhabditis chromosome arms are anchored to nuclear membrane via discontinuous association with LEM-2. Genome Biol. 2010;11:R120. doi: 10.1186/gb-2010-11-12-r120. PubMed DOI PMC

Mattout A., Pike B.L., Towbin B.D., Bank E.M., Gonzales-Sandoval A., Stadier M.B., Meister P., Gruenbaum Y., Gasser S.M. An EDMD mutation in C. elegans lamin blocks muscle-specific gene relocation and compromises muscle integrity. Curr. Biol. 2011;21:1603–1614. doi: 10.1016/j.cub.2011.08.030. PubMed DOI

Towbin B.D., Gonzáles-Aguilera C., Sack R., Gaidatzis D., Kalck V., Meister P., Askjaer P., Gasser S.M. Step-wise methylation of histone H3K9 positions heterochromatin at the nuclear periphery. Cell. 2012;150:934–947. doi: 10.1016/j.cell.2012.06.051. PubMed DOI

Campisi J., d’Adda di Fagagna F. Cellular senescence: When bad things happen to good cells. Nat. Rev. Mol. Cell Biol. 2007;8:729–740. doi: 10.1038/nrm2233. PubMed DOI

Narita M., Nunez S., Heard E., Narita M., Lin A.W., Hearn S.A., Spector D.L., Hannon G.J., Lowe S.W. Rb-mediated heterochromatin formation and silencing of E2F target genes during cellular senescence. Cell. 2003;113:703–716. doi: 10.1016/S0092-8674(03)00401-X. PubMed DOI

Dimri G.P., Lee X., Basile G., Acosta M., Scott G., Roskelley C., Medrano E.E., Linskens M., Rubelj I., Pereira-Smith O. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl. Acad. Sci. USA. 1995;92:9363–9367. doi: 10.1073/pnas.92.20.9363. PubMed DOI PMC

Kuilman T., Michaloglou C., Vredeveld L.C., Douma S., van Doorn R., Desmet C.J., Aarden L.A., Mooi W.J., Peeper D.S. Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell. 2008;133:1019–1031. doi: 10.1016/j.cell.2008.03.039. PubMed DOI

Novakova Z., Hubackova S., Kosar M., Janderova-Rossmeislova L., Dobrovolna J., Vasicova P., Vancurova M., Horejsi Z., Hozak P., Bartek J., et al. Cytokine expression and signaling in drug-induced cellular senescence. Oncogene. 2010;29:273–284. doi: 10.1038/onc.2009.318. PubMed DOI

d’Adda di Fagagna F., Reaper P.M., Clay-Farrace L., Fiegler H., Carr P., Von Zglinicki T., Saretzki G., Carter N.P., Jackson S.P. A DNA damage checkpoint response in telomere-initiated senescence. Nature. 2003;426:194–198. doi: 10.1038/nature02118. PubMed DOI

Von Zglinicki T., Saretzki G., Ladhoff J., d’Adda di Fagagna F., Jackson S.P. Human cell senescence as a DNA damage response. Mech. Ageing Dev. 2005;126:111–117. doi: 10.1016/j.mad.2004.09.034. PubMed DOI

Sedivy J.M. Telomeres limit cancer growth by inducing senescence: Long-sought in vivo evidence obtained. Cancer Cell. 2007;11:389–391. doi: 10.1016/j.ccr.2007.04.014. PubMed DOI

Von Zglinicki T. Oxidative stress shortens telomeres. Trends Biochem. Sci. 2002;27:339–344. doi: 10.1016/S0968-0004(02)02110-2. PubMed DOI

Serrano M., Lin A.W., McCurrach M.E., Beach D., Lowe S.W. Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell. 1997;88:593–602. doi: 10.1016/S0092-8674(00)81902-9. PubMed DOI

Kosar M., Bartkova J., Hubackova S., Hodny Z., Lukas J., Bartek J. Senescence-asociated heterochromatin foci are dispensable for cellular senescence, occur in a cell type- and insult-dependent manner and follow expression of p16ink4a. Cell Cycle. 2011;10:459–468. doi: 10.4161/cc.10.3.14707. PubMed DOI

Chen Q.M., Bartholomew J.C., Campisi J., Acosta M., Reagan J.D., Ames B.N. Molecular analysis of H2O2-induced senescent-like growth arrest in normal human fibroblasts: P53 and Rb control G1 arrest but not cellreplication. Biochem. J. 1998;332:43–50. doi: 10.1042/bj3320043. PubMed DOI PMC

Beausejour C.M., Krtolica A., Galimi F., Narita M., Lowe S.W., Yaswen P., Campisi J. Reversal of human cellular senescence: Roles of the p53 and p16 pathways. EMBO J. 2003;22:4212–4222. doi: 10.1093/emboj/cdg417. PubMed DOI PMC

Michaloglou C., Vredeveld L.C.W., Soengas M.S., Denoyelle C., Kuilman T., van der Horst C.M., Majoor D.M., Shay J.W., Mooi W.J., Peeper D.S. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature. 2005;436:720–724. doi: 10.1038/nature03890. PubMed DOI

Bartkova J., Rezaei N., Lintos M., Karakaidos P., Kletsas D., Issaeva N., Vassiliou L.F., Kolettas E., Niforou K., Zoumpourlis V.C., et al. Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature. 2006;444:633–637. doi: 10.1038/nature05268. PubMed DOI

Lukášová E., Kovařík A., Bačíková A., Falk M., Kozubek S. Loss of lamin B receptor is necessary to induce cellular senescence. Biochem. J. 2017;474:281–290. doi: 10.1042/BCJ20160459. PubMed DOI

Zhang R., Chen W., Adams P.D. Molecular dissection of formation of senescence-associated heterochromati foci. Mol. Cell. Biol. 2007;27:2343–2358. doi: 10.1128/MCB.02019-06. PubMed DOI PMC

Shimi T., Butin-Israeli V., Adam S.A., Hamanaka R.B., Goldman A.E., Lucas C.A., Shumaker D.K., Kosak S.T., Chandel N.S., Goldman R.D. The role of lamin B1 in cell proliferation and senescence. Genes Dev. 2011;25:2579–2593. doi: 10.1101/gad.179515.111. PubMed DOI PMC

Freund A., Laberge R.M., Demaria M., Campisi J. Lamin B1 loss is a senescence-associated biomarker. Mol. Biol. Cell. 2012;23:2066–2074. doi: 10.1091/mbc.E11-10-0884. PubMed DOI PMC

Dreesen O., Chojnowski A., Ong P.F., Zhao T.Y., Common J.E., Lunny D., Lane E.B., Lee S.J., Vardy L.A., Stewart C.L., et al. Lamin B1 fluctuations have differential effects on cellular proliferation and senescence. J. Cell Biol. 2013;200:605–617. doi: 10.1083/jcb.201206121. PubMed DOI PMC

Shah P.P., Donahue G., Otte G.L., Capell B.C., Nelson D.M., Cao K., Aggarwala V., Cruickshanks H.A., Rai T.S., McBryan T., et al. Lamin B1 depletion in senescence cells triggers large-scale changes in gene expression and the chromatin landscape. Genes Dev. 2013;27:1787–1799. doi: 10.1101/gad.223834.113. PubMed DOI PMC

Chandra T., Ewels P.A., Schoenfelder S., Furlan-Magaril M., Wingett S.W., Kirschner K., Thuret J.-Y., Andrews S., Fraser P., Reik W. Global reorganization of the nuclear landscape in senescence cells. Cell Rep. 2015;10:471–483. doi: 10.1016/j.celrep.2014.12.055. PubMed DOI PMC

Chandra T., Kirschner K., Thuret J.-Y., Pope B.J., Ryba T., Newman S., Ahmed K., Samarajiwa S.A., Salama R., Carroll T., et al. Independence of repressive histone markers and chromatin compaction during senescent heterochromatic layer formation. Mol. Cell. 2012;47:203–214. doi: 10.1016/j.molcel.2012.06.010. PubMed DOI PMC

Sadaie M., Salama R., Carroll T., Tomimatsu K., Chandra T., Young A.R.J., Narita M., Pérez-Mancera P.A., Bennett D.C., Chong H., et al. Redistribution of the lamin B1 genomic binding profile affects rearrangement of heterochromatic domains and SAHF formation during senescence. Genes Dev. 2015;27:1800–1813. doi: 10.1101/gad.217281.113. PubMed DOI PMC

Ye Q., Worman H.J. Primary structure analysis and lamin B and DNA binding of human LBR, an integral protein of the nuclear envelope inner membrane. J. Biol. Chem. 1994;269:11306–11311. PubMed

Worman H.J., Yuan J., Blobel G., Georgatos S.P. A lamin B receptor in the nuclear envelope. Proc. Natl. Acad. Sci. USA. 1988;85:8531–8534. doi: 10.1073/pnas.85.22.8531. PubMed DOI PMC

Ye Q., Worman H.J. Interaction between an integral protein of the nuclear envelope inner membrane and human chromodomain proteins to Drosophilla HP1. J. Biol. Chem. 1996;271:14653–14656. doi: 10.1074/jbc.271.25.14653. PubMed DOI

Von Mikecz A., Chen M., Rockel T., Scharf A. The nuclear ubiquitin-proteasome system: Visualization of proteasomes, protein aggregates, and proteolysis in the cell nucleus. Nucleus. 2008;463:191–202. PubMed

De Cecco M., Criscione S.W., Peckham E.J., Hillenmeyer S., Hamm E.A., Manivannan J., Peterson A.L., Kreiling J.A., Neretti N., Sedivy J.M. Genomes of replicative senescent cells undergo global epigenetic changes leading to gene silencing and activation of transposable elements. Aging Cell. 2013;12:247–256. doi: 10.1111/acel.12047. PubMed DOI PMC

Malhas A., Lee C.F., Sanders R., Sounders N.J., Vaux D.J. Defects in lamin B1 expression or processing affect interphase chromosome position and gene expression. J. Cell Biol. 2007;176:593–603. doi: 10.1083/jcb.200607054. PubMed DOI PMC

Nikolakaki E., Meier J., Simons G., Georgatos S.D., Giannakouros T. Mitotic phosphorylation of the lamin B receptor by a serine/arginine kinase and p34(cdc2) J. Biol. Chem. 1997;272:6208–6213. doi: 10.1074/jbc.272.10.6208. PubMed DOI

Duband-Goulet I., Courvalin J.-C., Buendia B. LBR, a chromatin and lamin binding protein from the inner nuclear membrane, is proteolysed at late stage of apoptosis. J. Cell Sci. 1998;111:1441–1451. PubMed

Ellenberg J., Siggia E.D., Moreira J.E., Smith C.L., Presley J.F., Worman H.J., Lippincott-Schwartz J. Nuclear membrane dynamics and reassembly in living cells: Targeting of an inner nuclear membrane protein in interphase and mitosis. J. Cell Biol. 1997;138:1193–1206. doi: 10.1083/jcb.138.6.1193. PubMed DOI PMC

Nikolakaki E., Milonis I., Giannakouros T. Lamin B receptor: Interplay between structure, function and localization. Cells. 2017;6:28. doi: 10.3390/cells6030028. PubMed DOI PMC

Tseng L.C., Chen R.H. Temporal control of nuclear envelope assembly by phosphorylation of lamin B receptor. Mol. Biol. Cell. 2011;22:3306–3317. doi: 10.1091/mbc.E11-03-0199. PubMed DOI PMC

Ivanov A., Pawlowski J., Manoharan I., van Tuyn J., Nelson D.M., Rai T.S., Shah P.S., Hewitt G., Korolchuk V.I., Passos J.F., et al. Lysozome-mediated processing of chromatin in senescence. J. Cell Biol. 2012;202:129–143. doi: 10.1083/jcb.201212110. PubMed DOI PMC

Peric-Hupkes D., Meuleman W., Pagie L., Bruggeman S.W., Solovei I., Brugman W., Graf S., Flicek P., Kerkhoven R.M., Reinders M., et al. Molecular maps of the reorganization of genom-nuclear lamina interactions during differentiation. Mol. Cell. 2010;38:603–613. doi: 10.1016/j.molcel.2010.03.016. PubMed DOI PMC

Meulman W., Peric-Hupkes D., Kind J., Beaudry J.B., Pagie L., Kellis M., Reinders M., Wessels L., van Steensel B. Constitutive nuclear lamina-genomeinteractions are highly conserved and associated with A/T-rich sequence. Genome Res. 2013;23:270–280. doi: 10.1101/gr.141028.112. PubMed DOI PMC

Solovei I., Thanisch K., Feodorova Y. How to rule the nucleus: Divide et impera. Curr. Opin. Cell Biol. 2016;40:47–59. doi: 10.1016/j.ceb.2016.02.014. PubMed DOI

Wijchers P.J., Geeven G., Eyres M., Bergsma A.J., Janssen M., Versteegen M., Zhu Y., Schell Y., Vermeulen C., de Vit E., et al. Characterization and dynamics of pericentromere-associated domains in mice. Genome Res. 2015;25:958–969. doi: 10.1101/gr.186643.114. PubMed DOI PMC

Solovei I., Schemelleh L., During K., Engelhardt A., Stein S., Cremer C., Cremer T. Differences in centromere positioning of cycling and postmitotic human cell types. Chromosoma. 2004;112:410–423. doi: 10.1007/s00412-004-0287-3. PubMed DOI

Bouwman B.A., de Laat W. Getting the genome in shape: The formation of loops, domains and compartments. Genome Biol. 2015;16:154. doi: 10.1186/s13059-015-0730-1. PubMed DOI PMC

Weierich C., Brero A., Stein S., von Hase J., Cremer C., Cremer T., Solovei I. Three-dimensional arrangements of centromeres and telomeres in nuclei of human and murine lymphocytes. Chromosom. Res. 2003;11:485–502. doi: 10.1023/A:1025016828544. PubMed DOI

Swanson E.C., Manning B., Zhang H., Lawrence J.B. Higher-order unfolding of satellite heterochromatin is a consistent and early event in cell senescence. J. Cell Biol. 2013;203:929–942. doi: 10.1083/jcb.201306073. PubMed DOI PMC