DERP6 (ELP5) and C3ORF75 (ELP6) regulate tumorigenicity and migration of melanoma cells as subunits of Elongator

[en] The Elongator complex is composed of 6 subunits (Elp1-Elp6) and promotes RNAPII transcript elongation through histone acetylation in the nucleus as well as tRNA modification in the cytoplasm. This acetyltransferase complex directly or indirectly regulates numerous biological processes ranging from exocytosis and resistance to heat shock in yeast to cell migration and neuronal differentiation in higher eukaryotes. The identity of human ELP1 through ELP4 has been reported but human ELP5 and ELP6 have remained uncharacterized. Here, we report that DERP6 (ELP5) and C3ORF75 (ELP6) encode these subunits of human Elongator. We further investigated the importance and function of these two subunits by a combination of biochemical analysis and cellular assays. Our results show that DERP6/ELP5 is required for the integrity of Elongator and directly connects ELP3 to ELP4. Importantly, the migration and tumorigenicity of melanomaderived cells are significantly decreased upon Elongator depletion through ELP1 or ELP3. Strikingly, DERP6/ELP5 and C3ORF75/ELP6-depleted melanoma cells have similar defects, further supporting the idea that DERP6/ELP5 and C3ORF75/ELP6 are essential for Elongator function. Together, our data identify DERP6/ELP5 and C3ORF75/ELP6 as key players for migration, invasion and tumorigenicity of melanoma cells, as integral subunits of Elongator.

Research Center/Unit :

Giga-Signal Transduction - ULiège

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Close, Pierre ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale

Gillard, Magali

Ladang, Aurélie ; Université de Liège - ULiège > GIGA - Membres

Jiang, Zheshen ; Université de Liège - ULiège > GIGA - Membres

Papuga, Jessica ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale

Hawkes, Nicola

Nguyen, Laurent ; Université de Liège - ULiège > Département des sciences cliniques > Neurologie

CHAPELLE, Jean-Paul ; Centre Hospitalier Universitaire de Liège - CHU > Chimie médicale

Bouillenne, Fabrice ; Université de Liège - ULiège > Centre d'ingénierie des protéines

Svejstrup, Jesper

Fillet, Marianne ; Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments

Chariot, Alain ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale

Language :

English

Title :

DERP6 (ELP5) and C3ORF75 (ELP6) regulate tumorigenicity and migration of melanoma cells as subunits of Elongator

Publication date :

01 August 2012

Journal title :

Journal of Biological Chemistry

ISSN :

0021-9258

eISSN :

1083-351X

Publisher :

American Society for Biochemistry and Molecular Biology, Baltimore, United States - Maryland

Peer reviewed :

Peer Reviewed verified by ORBi

Additional URL :

http://www.jbc.org/content/early/2012/08/01/jbc.M112.402727

Available on ORBi :

since 03 August 2012

Statistics

Number of views

105 (23 by ULiège)

Number of downloads

7 (7 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

OpenAlex citations

Bibliography

Otero, G., Fellows, J., Li, Y., de Bizemont, T., Dirac, A. M., Gustafsson, C. M., Erdjument-Bromage, H., Tempst, P., and Svejstrup, J. Q. (1999) Elongator, a multisubunit component of a novel RNA polymerase II holoenzyme for transcriptional elongation. Mol. Cell 3, 109-118 (Pubitemid 29292482)
Kim, J. H., Lane, W. S., and Reinberg, D. (2002) Human Elongator facilitates RNA polymerase II transcription through chromatin. Proc. Natl. Acad. Sci. U.S.A. 99, 1241-1246 (Pubitemid 34136477)
Hawkes, N. A., Otero, G., Winkler, G. S., Marshall, N., Dahmus, M. E., Krappmann, D., Scheidereit, C., Thomas, C. L., Schiavo, G., Erdjument-Bromage, H., Tempst, P., and Svejstrup, J. Q. (2002) Purification and characterization of the human elongator complex. J. Biol. Chem. 277, 3047-3052 (Pubitemid 34953342)
Krogan, N. J., and Greenblatt, J. F. (2001) Characterization of a six-subunit holo-elongator complex required for the regulated expression of a group of genes in Saccharomyces cerevisiae. Mol. Cell Biol. 21, 8203-8212 (Pubitemid 33051809)
Wittschieben, B. O., Otero, G., de Bizemont, T., Fellows, J., Erdjument-Bromage, H., Ohba, R., Li, Y., Allis, C. D., Tempst, P., and Svejstrup, J. Q. (1999) A novel histone acetyltransferase is an integral subunit of elongating RNA polymerase II holoenzyme. Mol. Cell 4, 123-128 (Pubitemid 29386138)
Winkler, G. S., Kristjuhan, A., Erdjument-Bromage, H., Tempst, P., and Svejstrup, J. Q. (2002) Elongator is a histone H3 and H4 acetyltransferase important for normal histone acetylation levels in vivo. Proc. Natl. Acad. Sci. U.S.A. 99, 3517-3522 (Pubitemid 34252140)
Kristjuhan, A., Walker, J., Suka, N., Grunstein, M., Roberts, D., Cairns, B. R., and Svejstrup, J. Q. (2002) Transcriptional inhibition of genes with severe histone h3 hypoacetylation in the coding region. Mol. Cell 10, 925-933 (Pubitemid 35335646)
Gilbert, C., Kristjuhan, A., Winkler, G. S., and Svejstrup, J. Q. (2004) Elongator interactions with nascent mRNA revealed by RNA immunoprecipitation. Mol. Cell 14, 457-464 (Pubitemid 38648799)
Métivier, R., Penot, G., Hübner, M. R., Reid, G., Brand, H., Kos, M., and Gannon, F. (2003) Estrogen receptor-α directs ordered, cyclical, and combinatorial recruitment of cofactors on a natural target promoter. Cell 115, 751-763 (Pubitemid 38030303)
Kouskouti, A., and Talianidis, I. (2005) Histone modifications defining active genes persist after transcriptional and mitotic inactivation.EMBOJ. 24, 347-357 (Pubitemid 40216139)
Close, P., Hawkes, N., Cornez, I., Creppe, C., Lambert, C. A., Rogister, B., Siebenlist, U., Merville, M. P., Slaugenhaupt, S. A., Bours, V., Svejstrup, J. Q., and Chariot, A. (2006) Transcription impairment and cell migration defects in elongator-depleted cells: implication for familial dysautonomia. Mol. Cell 22, 521-531 (Pubitemid 44255542)
Li, Y., Takagi, Y., Jiang, Y., Tokunaga, M., Erdjument-Bromage, H., Tempst, P., and Kornberg, R. D. (2001) A multiprotein complex that interacts with RNA polymerase II elongator. J. Biol. Chem. 276, 29628-29631
Holmberg, C., Katz, S., Lerdrup, M., Herdegen, T., Jää ttelä, M., Aronheim, A., and Kallunki, T. (2002) A novel specific role for I κB kinase complex-associated protein in cytosolic stress signaling. J. Biol. Chem. 277, 31918-31928 (Pubitemid 34969000)
Svejstrup, J. Q. (2007) Elongator complex: how many roles does it play? Curr. Opin. Cell Biol. 19, 331-336
Huang, B., Johansson, M. J., and Byström, A. S. (2005) An early step in wobble uridine tRNA modification requires the Elongator complex. RNA 11, 424-436 (Pubitemid 40397176)
Esberg, A., Huang, B., Johansson, M. J., and Byström, A. S. (2006) Elevated levels of two tRNA species bypass the requirement for elongator complex in transcription and exocytosis. Mol. Cell 24, 139-148 (Pubitemid 44466676)
Bauer, F., Matsuyama, A., Candiracci, J., Dieu, M., Scheliga, J., Wolf, D. A., Yoshida, M., and Hermand, D. (2012) Translational control of cell division by elongator. Cell Rep. 1, 424-433
Mehlgarten, C., Jablonowski, D., Wrackmeyer, U., Tschitschmann, S., Sondermann, D., Jäger, G., Gong, Z., Byström, A. S., Schaffrath, R., and Breunig, K. D. (2010) Elongator function in tRNA wobble uridine modification is conserved between yeast and plants. Mol. Microbiol. 76, 1082-1094
Chen, C., Tuck, S., and Byström, A. S. (2009) Defects in tRNA modification associated with neurological and developmental dysfunctions in Caenorhabditis elegans elongator mutants. PLoS genetics 5, e1000561
Versées, W., De Groeve, S., and Van Lijsebettens, M. (2010) Elongator, a conserved multitasking complex? Mol. Microbiol. 76, 1065-1069
Creppe, C., Malinouskaya, L., Volvert, M. L., Gillard, M., Close, P., Malaise, O., Laguesse, S., Cornez, I., Rahmouni, S., Ormenese, S., Belachew, S., Malgrange, B., Chapelle, J. P., Siebenlist, U., Moonen, G., Chariot, A., and Nguyen, L. (2009) Elongator controls the migration and differentiation of cortical neurons through acetylation of α-tubulin. Cell 136, 551-564
Solinger, J. A., Paolinelli, R., Klöss, H., Scorza, F. B., Marchesi, S., Sauder, U., Mitsushima, D., Capuani, F., Stärzenbaum, S. R., and Cassata, G. (2010) The Caenorhabditis elegans Elongator complex regulates neuronal α-tubulin acetylation. PLoS genetics 6, e1000820
Miskiewicz, K., Jose, L. E., Bento-Abreu, A., Fislage, M., Taes, I., Kasprowicz, J., Swerts, J., Sigrist, S., Versées, W., Robberecht, W., and Verstreken, P. (2011) ELP3 controls active zone morphology by acetylating the ELKS family member Bruchpilot. Neuron 72, 776-788
Rahl, P. B., Chen, C. Z., and Collins, R. N. (2005) Elp1p, the yeast homolog of the FD disease syndrome protein, negatively regulates exocytosis independently of transcriptional elongation. Mol. Cell 17, 841-853 (Pubitemid 40386945)
Chen, C., Huang, B., Eliasson, M., Rydén, P., and Byström, A. S. (2011) Elongator complex influences telomeric gene silencing and DNA damage response by its role in wobble uridine tRNA modification. PLoS genetics 7, e1002258
Li, Q., Fazly, A. M., Zhou, H., Huang, S., Zhang, Z., and Stillman, B. (2009) The elongator complex interacts with PCNA and modulates transcriptional silencing and sensitivity to DNA damage agents. PLoS genetics 5, e1000684
Nelissen, H., Fleury, D., Bruno, L., Robles, P., De Veylder, L., Traas, J., Micol, J. L., Van Montagu, M., Inzé, D., and Van Lijsebettens, M. (2005) The elongata mutants identify a functional Elongator complex in plants with a role in cell proliferation during organ growth. Proc. Natl. Acad. Sci. U.S.A. 102, 7754-7759 (Pubitemid 40744244)
Walker, J., Kwon, S. Y., Badenhorst, P., East, P., McNeill, H., and Svejstrup, J. Q. (2011) Role of elongator subunit Elp3 in Drosophila melanogaster larval development and immunity. Genetics 187, 1067-1075
Chen, Y. T., Hims, M. M., Shetty, R. S., Mull, J., Liu, L., Leyne, M., and Slaugenhaupt, S. A. (2009) Loss of mouse Ikbkap, a subunit of elongator, leads to transcriptional deficits and embryonic lethality that can be rescued by human IKBKAP. Mol. Cell Biol. 29, 736-744
Slaugenhaupt, S. A., and Gusella, J. F. (2002) Familial dysautonomia. Curr. Opin. Genet. Dev. 12, 307-311 (Pubitemid 34663764)
Gold-von Simson, G., and Axelrod, F. B. (2006) Familial dysautonomia: update and recent advances. Curr. Probl. Pediatr Adolesc Health Care 36, 218-237 (Pubitemid 43843082)
Anderson, S. L., Coli, R., Daly, I. W., Kichula, E. A., Rork, M. J., Volpi, S. A., Ekstein, J., and Rubin, B. Y. (2001) Familial dysautonomia is caused by mutations of the IKAP gene. Am. J. Hum. Genet. 68, 753-758 (Pubitemid 32202767)
Slaugenhaupt, S. A., Blumenfeld, A., Gill, S. P., Leyne, M., Mull, J., Cuajungco, M. P., Liebert, C. B., Chadwick, B., Idelson, M., Reznik, L., Robbins, C., Makalowska, I., Brownstein, M., Krappmann, D., Scheidereit, C., Maayan, C., Axelrod, F. B., and Gusella, J. F. (2001) Tissue-specific expression of a splicing mutation in the IKBKAP gene causes familial dysautonomia. Am. J. Hum. Genet. 68, 598-605 (Pubitemid 32202755)
Simpson, C. L., Lemmens, R., Miskiewicz, K., Broom, W. J., Hansen, V. K., van Vught, P. W., Landers, J. E., Sapp, P., Van Den Bosch, L., Knight, J., Neale, B. M., Turner, M. R., Veldink, J. H., Ophoff, R. A., Tripathi, V. B., Beleza, A., Shah, M. N., Proitsi, P., Van Hoecke, A., Carmeliet, P., Horvitz, H. R., Leigh, P. N., Shaw, C. E., van den Berg, L. H., Sham, P. C., Powell, J. F., Verstreken, P., Brown, R. H., Jr., Robberecht, W., and Al-Chalabi, A. (2009) Variants of the elongator protein 3 (ELP3) gene are associated with motor neuron degeneration. Hum. Mol. Genet. 18, 472-481
Nguyen, L., Humbert, S., Saudou, F., and Chariot, A. (2010) Elongator-an emerging role in neurological disorders. Trends Mol. Med. 16, 1-6
Johansen, L. D., Naumanen, T., Knudsen, A., Westerlund, N., Gromova, I., Junttila, M., Nielsen, C., Bøttzauw, T., Tolkovsky, A., Westermarck, J., Coffey, E. T., Jäättelä, M., and Kallunki, T. (2008) IKAP localizes to membrane ruffles with filamin A and regulates actin cytoskeleton organization and cell migration. J. Cell Sci. 121, 854-864 (Pubitemid 351578121)
Boone, N., Loriod, B., Bergon, A., Sbai, O., Formisano-Tréziny, C., Gabert, J., Khrestchatisky, M., Nguyen, C., Féron, F., Axelrod, F. B., and Ibrahim, E. C. (2010) Olfactory stem cells, a new cellular model for studying molecular mechanisms underlying familial dysautonomia. PLoS One 5, e15590
Lee, G., Papapetrou, E. P., Kim, H., Chambers, S. M., Tomishima, M. J., Fasano, C. A., Ganat, Y. M., Menon, J., Shimizu, F., Viale, A., Tabar, V., Sadelain, M., and Studer, L. (2009) Modelling pathogenesis and treatment of familial dysautonomia using patient-specific iPSCs. Nature 461, 402-406
Petrakis, T. G., Wittschieben, B. Ø., and Svejstrup, J. Q. (2004) Molecular architecture, structure-function relationship, and importance of the Elp3 subunit for the RNA binding of holo-elongator. J. Biol. Chem. 279, 32087-32092 (Pubitemid 39014654)
Viatour, P., Dejardin, E., Warnier, M., Lair, F., Claudio, E., Bureau, F., Marine, J. C., Merville, M. P., Maurer, U., Green, D., Piette, J., Siebenlist, U., Bours, V., and Chariot, A. (2004) GSK3-mediated BCL-3 phosphorylation modulates its degradation and its oncogenicity. Mol. Cell 16, 35-45 (Pubitemid 39330150)
Close, P., East, P., Dirac-Svejstrup, A. B., Hartmann, H., Heron, M., Maslen, S., Chariot, A., Söding, J., Skehel, M., and Svejstrup, J. Q. (2012) DBIRD complex integrates alternative mRNA splicing with RNA polymerase II transcript elongation. Nature 484, 386-389
Gobeil, S., Zhu, X., Doillon, C. J., and Green, M. R. (2008) A genome-wide shRNA screen identifies GAS1 as a novel melanoma metastasis suppressor gene. Genes Dev. 22, 2932-2940
Sebastiaan Winkler, G., Lacomis, L., Philip, J., Erdjument-Bromage, H., Svejstrup, J. Q., and Tempst, P. (2002) Isolation and mass spectrometry of transcription factor complexes. Methods 26, 260-269 (Pubitemid 34975669)
Yuan, J., Tang, W., Luo, K., Chen, X., Gu, X., Wan, B., and Yu, L. (2006) Cloning and characterization of the human gene DERP6, which activates transcriptional activities of p53. Molecular Biol. Rep. 33, 151-158 (Pubitemid 44087086)
Winkler, G. S., Petrakis, T. G., Ethelberg, S., Tokunaga, M., Erdjument-Bromage, H., Tempst, P., and Svejstrup, J. Q. (2001) RNA polymerase II elongator holoenzyme is composed of two discrete subcomplexes. J. Biol. Chem. 276, 32743-32749
Pokholok, D. K., Hannett, N. M., and Young, R. A. (2002) Exchange of RNA polymerase II initiation and elongation factors during gene expression in vivo. Mol Cell 9, 799-809 (Pubitemid 34454890)
Doillon, C. J., Gagnon, E., Paradis, R., and Koutsilieris, M. (2004) Threedimensional culture system as a model for studying cancer cell invasion capacity and anticancer drug sensitivity. Anticancer Res. 24, 2169-2177 (Pubitemid 39166291)
Glatt, S., Létoquart, J., Faux, C., Taylor, N. M., Séraphin, B., and Müller, C. W. (2012) The Elongator subcomplex Elp456 is a hexameric RecA-like ATPase. Nat. Struct. Mol. Biol. 19, 314-320
Lin, Z., Zhao, W., Diao, W., Xie, X., Wang, Z., Zhang, J., Shen, Y., and Long, J. (2012) Crystal structure of elongator subcomplex elp4-6. J. Biol. Chem. 287, 21501-21508
Fellows, J., Erdjument-Bromage, H., Tempst, P., and Svejstrup, J. Q. (2000) The Elp2 subunit of elongator and elongating RNA polymerase II holoenzyme is a WD40 repeat protein. J. Biol. Chem. 275, 12896-12899 (Pubitemid 30241445)
Greenwood, C., Selth, L. A., Dirac-Svejstrup, A. B., and Svejstrup, J. Q. (2009) An iron-sulfur cluster domain in Elp3 important for the structural integrity of elongator. J. Biol. Chem. 284, 141-149
Cornez, I., Creppe, C., Gillard, M., Hennuy, B., Chapelle, J. P., Dejardin, E., Merville, M. P., Close, P., and Chariot, A. (2008) Deregulated expression of pro-survival and pro-apoptotic p53-dependent genes upon Elongator deficiency in colon cancer cells. Biochem. Pharmacol. 75, 2122-2134
Bolukbasi, E., Vass, S., Cobbe, N., Nelson, B., Simossis, V., Dunbar, D. R., and Heck, M. M. (2012) Drosophila poly suggests a novel role for the Elongator complex in insulin receptor-target of rapamycin signalling. Open Biol. J. 2, 110031