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Intracellular trafficking of emerin, the Emery-Dreifuss muscular dystrophy protein
C. Ostlund, J. Ellenberg, E. Hallberg, J. Lippincott-Schwartz, H.J. Worman
Journal of Cell Science 1999 112: 1709-1719;

Summary

Emerin is an integral protein of the inner nuclear membrane that is mutated or not expressed in patients with Emery-Dreifuss muscular dystrophy. Confocal immunofluorescence microscopy studies of the intracellular targeting of truncated forms of emerin, some of which are found in patients with Emery-Dreifuss muscular dystrophy, show that the nucleoplasmic, amino-terminal domain is necessary and sufficient for nuclear retention. When this domain is fused to a transmembrane segment of an integral membrane protein of the ER/plasma membrane, the chimeric protein is localized in the inner nuclear membrane. The transmembrane segment of emerin is not targeted to the inner nuclear membrane. Fluorescence photobleaching experiments of emerin fused to green fluorescent protein demonstrate that the diffusional mobility (D) of emerin is decreased in the inner nuclear membrane (D=0.10+/-0.01 microm2/second) compared to the ER membrane (D=0.32+/-0.01 microm2/second). This is in agreement with a model where integral proteins reach the inner nuclear membrane by lateral diffusion and are retained there by association with nucleoplasmic components. Some overexpressed emerin-green fluorescent protein also reaches the plasma membrane of transfected cells, where its diffusion is similar to that in the inner nuclear membrane, suggesting that emerin may also associate with non-nuclear structures.

REFERENCES

    1. Abney J. R.,
    2. Cutler B.,
    3. Fillbach M. L.,
    4. Axelrod D. and
    5. Scalettar B. A.
    (1997). Chromatin dynamics in interphase nuclei and its implications for nuclear structure. J. Cell Biol 137, 1459–1468
    OpenUrlAbstract/FREE Full Text
    1. Adam S. A.,
    2. Sterne-Marr R. and
    3. Gerace L.
    (1992). Nuclear protein import using digitonin-permeabilized cells. Methods Enzymol 219, 97–110
    OpenUrlCrossRefPubMedWeb of Science
    1. Aebi U.,
    2. Cohn J.,
    3. Buhle L. and
    4. Gerace L.
    (1986). The nuclear lamina is a meshwork of intermediate-type filaments. Nature 323, 560–564
    OpenUrlCrossRefPubMed
    1. Bione S.,
    2. Maestrini E.,
    3. Rivella S.,
    4. Mancini M.,
    5. Regis S.,
    6. Romeo G. and
    7. Toniolo D.
    (1994). Identification of a novel X-linked gene responsible for Emery-Dreifuss muscular dystrophy. Nat. Genet 8, 323–327
    OpenUrlCrossRefPubMedWeb of Science
    1. Bione S.,
    2. Small K.,
    3. Aksmanovic V. M. A.,
    4. D'Urso M.,
    5. Ciccodicola A.,
    6. Merlini L.,
    7. Morandi L.,
    8. Kress W.,
    9. Yates J. R. W.,
    10. Warren S. T. and
    11. Toniolo D.
    (1995). Identification of new mutations in the Emery-Dreifuss muscular dystrophy gene and evidence for genetic heterogeneity of the disease. Hum. Mol. Genet 4, 1859–1863
    OpenUrlAbstract/FREE Full Text
    1. Bonne G.,
    2. Raffaele di Barletta M.,
    3. Varnous S.,
    4. Becane H.-M.,
    5. Hammouda E.-H.,
    6. Merlini L.,
    7. Muntoni F.,
    8. Greenberg C. R.,
    9. Gary F.,
    10. Urtizberea J.-A.,
    11. Duboc D.,
    12. Fardeau M.,
    13. Toniolo D. and
    14. Schwartz K.
    (1999). Mutations in the gene encoding lamin A/C cause autosomal dominant Emery-Dreifuss muscular dystrophy. Nat. Genet 21, 285–288
    OpenUrlCrossRefPubMedWeb of Science
    1. Cartegni L.,
    2. Raffaele di Barletta M.,
    3. Barresi R.,
    4. Squarzoni S.,
    5. Sabatelli P.,
    6. Maraldi N.,
    7. Mora M.,
    8. Di Blasi C.,
    9. Cornelio F.,
    10. Merlini L.,
    11. Villa A.,
    12. Cobianchi F. and
    13. Toniolo D.
    (1997). Heart-specific localization of emerin: new insights into Emery-Dreifuss muscular dystrophy. Hum. Mol. Genet 6, 2257–2264
    OpenUrlAbstract/FREE Full Text
    1. Chiacchia K. B. and
    2. Drickamer K.
    (1984). Direct evidence for the transmembrane orientation of the hepatic glycoprotein receptors. J. Biol. Chem 259, 15440–15446
    OpenUrlAbstract/FREE Full Text
    1. Cole N. B.,
    2. Smith C. L.,
    3. Sciaky N.,
    4. Terasaki M.,
    5. Edidin M. and
    6. Lippincott-Schwartz J.
    (1996). Diffusional mobility of Golgi proteins in membranes of living cells. Science 273, 797–801
    OpenUrlAbstract
    1. Eissenberg J. C.,
    2. James T. C.,
    3. Foster-Hartnett D. M.,
    4. Hartnett T.,
    5. Ngan V. and
    6. Elgin S. C.
    (1990). Mutation in a heterochromatin-specific chromosomal protein is associated with suppression of position-effect variegation in Drosophila melanogaster. Proc. Natl. Acad. Sci. USA 87, 9923–9927
    OpenUrlAbstract/FREE Full Text
    1. Ellenberg J.,
    2. Siggia E. D.,
    3. Moreira J. E.,
    4. Smith C. L.,
    5. Presley J. F.,
    6. Worman H. J. and
    7. Lippincott-Schwartz J.
    (1997). Nuclear membrane dynamics and reassembly in living cells: targeting of an inner nuclear membrane protein in interphase and mitosis. J. Cell Biol 138, 1193–1206
    OpenUrlAbstract/FREE Full Text
    1. Ellis J. A.,
    2. Craxton M.,
    3. Yates J. R. W. and
    4. Kendrick-Jones J.
    (1998). Aberrant intracellular targeting and cell cycle-dependent phosphorylation of emerin contribute to the Emery-Dreifuss muscular dystrophy phenotype. J. Cell Sci 111, 781–792
    OpenUrlAbstract/FREE Full Text
    1. Emery A. E. H.
    (1989). Emery-Dreifuss syndrome. J. Med. Genet 26, 637–641
    OpenUrlAbstract/FREE Full Text
    1. Emery A. E. H. and
    2. Dreifuss F. E.
    (1966). Unusual type of benign X-linked muscular dystrophy. J.Neurol. Neurosurg. Psychiatry 29, 338–342
    OpenUrlFREE Full Text
    1. Fisher D. Z.,
    2. Chaudhary N. and
    3. Blobel G.
    (1986). cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins. Proc. Natl. Acad. Sci. USA 83, 6450–6454
    OpenUrlAbstract/FREE Full Text
    1. Foisner R. and
    2. Gerace L.
    (1993). Integral membrane proteins of the nuclear envelope interact with lamins and chromosomes, and binding is modulated by mitotic phosphorylation. Cell 73, 1267–1279
    OpenUrlCrossRefPubMedWeb of Science
    1. Frangioni J. V. and
    2. Neel B. G.
    (1993). Use of a general purpose mammalian expression vector for studying intracellular protein targeting: identification of critical residues in the nuclear lamin A/C nuclear localization signal. J. Cell Sci 105, 481–488
    OpenUrlAbstract/FREE Full Text
    1. Furukawa K.,
    2. Fritze C. E. and
    3. Gerace L.
    (1998). The major nuclear envelope domain of LAP2 coincides with its lamin binding region but is distinct from its chromatin interaction domain. J. Biol. Chem 273, 4213–4219
    OpenUrlAbstract/FREE Full Text
    1. Furukawa K.,
    2. Pante N.,
    3. Aebi U. and
    4. Gerace L.
    (1995). Cloning of a cDNA for lamina-associated polypeptide 2 (LAP2) and identification of regions that specify targeting to the nuclear envelope. EMBO J 14, 1626–1636
    OpenUrlPubMedWeb of Science
    1. Gilchrist J. M. and
    2. Leshner R. T.
    (1986). Autosomal dominant humeroperoneal myopathy. Arch. Neurol 43, 734–735
    OpenUrlCrossRefPubMed
    1. Görlich D. and
    2. Mattaj I. W.
    (1996). Nucleocytoplasmic transport. Science 271, 1513–1518
    OpenUrlAbstract
    1. Harris C. A.,
    2. Andryuk P. J.,
    3. Cline S.,
    4. Chan H. K.,
    5. Natarajan A.,
    6. Siekierka J. J. and
    7. Goldstein G.
    (1994). Three distinct human thymopoietins are derived from alternatively spliced mRNAs. Proc. Natl. Acad. Sci. USA 91, 6238–6287
    OpenUrl
    1. Holmer L.,
    2. Pezhman A. and
    3. Worman H. J.
    (1998). The human LBR/sterol reductase mutigene family. Genomics 54, 469–476
    OpenUrlCrossRefPubMedWeb of Science
    1. Lippincott-Schwartz J.,
    2. Yuan L. C.,
    3. Bonifacino J. S. and
    4. Klausner R. D.
    (1989). Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane recycling from Golgi to ER. Cell 56, 801–813
    OpenUrlCrossRefPubMedWeb of Science
    1. Manilal S.,
    2. Nguyen thi Man ,
    3. Sewry C. A. and
    4. Morris G. E.
    (1996). The Emery-Dreifuss muscular dystrophy protein, emerin, is a nuclear membrane protein. Hum. Mol. Genet 5, 801–808
    OpenUrlAbstract/FREE Full Text
    1. Manilal S.,
    2. Sewry C. A.,
    3. Pereboev A.,
    4. Nguyen thi Man ,
    5. Gobbi P.,
    6. Hawkes S.,
    7. Love D. R. and
    8. Morris G. E.
    (1999). Distribution of emerin and lamins in the heart and implications for Emery-Dreifuss muscular dystrophy. Hum. Mol. Genet 8, 353–359
    OpenUrlAbstract/FREE Full Text
    1. Marshall W. F.,
    2. Straight A.,
    3. Marko J. F.,
    4. Swedlow J.,
    5. Dernburg A.,
    6. Belmont A.,
    7. Murray A. W.,
    8. Agard D. A. and
    9. Sedat J. W.
    (1997). Interphase chromosomes undergo constrained diffusional motion in living cells. Curr. Biol 7, 930–939
    OpenUrlCrossRefPubMedWeb of Science
    1. Martin L.,
    2. Crimaudo C. and
    3. Gerace L.
    (1995). cDNA cloning and characterization of lamina-associated polypeptide 1C (LAP1C), an integral protein of the inner nuclear membrane. J. Biol. Chem 270, 8822–8828
    OpenUrlAbstract/FREE Full Text
    1. McKeon F. D.,
    2. Kirschner M. W. and
    3. Caput D.
    (1986). Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins. Nature 319, 463–468
    OpenUrlCrossRefPubMed
    1. Mellow T. E.,
    2. Halberg D. and
    3. Drickamer K.
    (1988). Endocytosis of N-acetyl-glucosamine-containing glycoproteins by rat fibroblasts expressing a single species of chicken liver glycoprotein receptor. J. Biol. Chem 263, 5468–5473
    OpenUrlAbstract/FREE Full Text
    1. Miller R. G.,
    2. Layzer R. B.,
    3. Mellenthin M. A.,
    4. Golabi M.,
    5. Francoz R. A. and
    6. Mall J. C.
    (1985). Emery-Dreifuss muscular dystrophy with autosomal dominant transmission. Neurology 35, 1230–1233
    OpenUrlAbstract/FREE Full Text
    1. Misumi Y.,
    2. Miki K.,
    3. Takatsuki A.,
    4. Tamura G. and
    5. Ikehara Y.
    (1986). Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes. J. Biol. Chem 261, 11398–11403
    OpenUrlAbstract/FREE Full Text
    1. Nagano A.,
    2. Koga R.,
    3. Ogawa M.,
    4. Kurano Y.,
    5. Kawada J.,
    6. Okada R.,
    7. Hayashi Y. K.,
    8. Tsukahara T. and
    9. Arahata K.
    (1996). Emerin deficiency at the nuclear membrane in patients with Emery-Dreifuss muscular dystrophy. Nat. Genet 12, 254–259
    OpenUrlCrossRefPubMedWeb of Science
    1. Nakamura N.,
    2. Rabouille C.,
    3. Watson R.,
    4. Nilsson T.,
    5. Hui N.,
    6. Slusarewicz P.,
    7. Kreis T. E. and
    8. Warren G.
    (1995). Characterization of a cis-Golgi matrix protein, GM130. J. Cell Biol 131, 1715–1726
    OpenUrlAbstract/FREE Full Text
    1. Paine P. L.,
    2. Moore L. C. and
    3. Horowitz S.
    (1975). Nuclear envelope permeability. Nature 254, 109–114
    OpenUrlCrossRefPubMed
    1. Pemberton L. F.,
    2. Blobel G. and
    3. Rosenblum J. S.
    (1998). Transport routes through the nuclear pore complex. Curr. Opin. Cell Biol 10, 392–399
    OpenUrlCrossRefPubMedWeb of Science
    1. Rowland L. P.,
    2. Fetell M.,
    3. Olarte M.,
    4. Hays A.,
    5. Singh N. and
    6. Wanat F. E.
    (1979). Emery-Dreifuss muscular dystrophy. Ann. Neurol 5, 111–117
    OpenUrlCrossRefPubMedWeb of Science
    1. Saiki R. K.,
    2. Gelfand D. H.,
    3. Stoffel S.,
    4. Scharf S. J.,
    5. Higuchi R.,
    6. Horn G. T.,
    7. Mullis K. B. and
    8. Erlich H.
    (1987). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487–491
    OpenUrlCrossRef
    1. Senior A. and
    2. Gerace L.
    (1988). Integral membrane proteins specific to the inner nuclear membrane and associated with the nuclear lamina. J. Cell Biol 107, 2029–2036
    OpenUrlAbstract/FREE Full Text
    1. Schuler E.,
    2. Lin F. and
    3. Worman H. J.
    (1994). Characterization of the human gene encoding LBR, an integral protein of the nuclear envelope inner membrane. J. Biol. Chem 269, 11312–11317
    OpenUrlAbstract/FREE Full Text
    1. Sciaky N.,
    2. Presley J.,
    3. Smith C.,
    4. Zaal K. J. M.,
    5. Cole N.,
    6. Moreira J. E.,
    7. Terasaki M.,
    8. Siggia E. and
    9. Lippincott-Schwartz J.
    (1997). Golgi tubule traffic and the effects of brefeldin A visualized in living cells. J. Cell Biol 139, 1137–1155
    OpenUrlAbstract/FREE Full Text
    1. Smith S. and
    2. Blobel G.
    (1993). The first membrane spanning region of the lamin B receptor is sufficient for sorting to the inner nuclear membrane. J. Cell Biol 120, 631–637
    OpenUrlAbstract/FREE Full Text
    1. Söderqvist H.,
    2. Imreh G.,
    3. Kihlmark M.,
    4. Linnman C.,
    5. Ringertz N. and
    6. Hallberg E.
    (1997). Intracellular distribution of an integral nuclear pore membrane protein fused to green fluorescent protein. Localization of a targeting domain. Eur. J. Biochem 250, 808–813
    OpenUrlPubMed
    1. Soullam B. and
    2. Worman H. J.
    (1993). The amino-terminal domain of the lamin B receptor is a nuclear envelope targeting signal. J. Cell Biol 120, 1093–1100
    OpenUrlAbstract/FREE Full Text
    1. Soullam B. and
    2. Worman H. J.
    (1995). Signals and structural features involved in integral membrane protein targeting to the inner nuclear membrane. J. Cell Biol 130, 15–27
    OpenUrlAbstract/FREE Full Text
    1. Squarzoni S.,
    2. Sabatelli P.,
    3. Ognibene A.,
    4. Toniolo D.,
    5. Cartegni L.,
    6. Cobianchi F.,
    7. Petrini S.,
    8. Merlini L. and
    9. Maraldi N. M.
    (1998). Immunocytochemical detection of emerin within the nuclear matrix. Neuromusc. Disord 8, 338–344
    OpenUrlCrossRefPubMed
    1. Worman H. J.,
    2. Evans C. D. and
    3. Blobel G.
    (1990). The lamin B receptor of the nuclear envelope inner membrane: a polytopic protein with eight potential transmembrane domains. J. Cell Biol 111, 1535–1542
    OpenUrlAbstract/FREE Full Text
    1. Worman H. J.,
    2. Yuan J.,
    3. Blobel G. and
    4. Georgatos S. D.
    (1988). A lamin B receptor in the nuclear envelope. Proc. Natl. Acad. Sci. USA 85, 8531–8534
    OpenUrlAbstract/FREE Full Text
    1. Wozniak R. W. and
    2. Blobel G.
    (1992). The single transmembrane segment of gp210 is sufficient for sorting to the pore membrane domain of the nuclear envelope. J. Cell Biol 119, 1441–1449
    OpenUrlAbstract/FREE Full Text
    1. Yates J. R. W.
    (1997). 43rd ENMC international workshop on Emery-Dreifuss muscular dystrophy, 22 June 1996, Naarden, the Netherlands. Neuromusc. Disord 7, 67–69
    OpenUrlCrossRefPubMed
    1. Ye. Q.,
    2. Callebaut I.,
    3. Pezhman A.,
    4. Courvalin J.-C. and
    5. Worman H. J.
    (1997). Domain-specific interactions of human HP1-type chromodomain proteins and inner nuclear membrane protein LBR. J. Biol. Chem 272, 14983–14989
    OpenUrlAbstract/FREE Full Text
    1. Ye Q. and
    2. Worman H. J.
    (1994). Primary structure analysis and lamin B and DNA binding of human LBR, an integral protein of the nuclear envelope inner membrane. J. Biol. Chem 269, 11306–11311
    OpenUrlAbstract/FREE Full Text
    1. Ye Q. and
    2. Worman H. J.
    (1996). Interaction between an integral protein of the nuclear envelope inner membrane and human chromodomain proteins homologous to Drosophila HP1. J. Biol. Chem 271, 14653–14656
    OpenUrlAbstract/FREE Full Text
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Journal Articles
Intracellular trafficking of emerin, the Emery-Dreifuss muscular dystrophy protein
C. Ostlund, J. Ellenberg, E. Hallberg, J. Lippincott-Schwartz, H.J. Worman
Journal of Cell Science 1999 112: 1709-1719;
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