Skip to main content
Top
Published in: Cellular Oncology 4/2011

01-08-2011 | Original Paper

Nuclear localization of the mitochondrial ncRNAs in normal and cancer cells

Authors: Eduardo Landerer, Jaime Villegas, Veronica A. Burzio, Luciana Oliveira, Claudio Villota, Constanza Lopez, Franko Restovic, Ronny Martinez, Octavio Castillo, Luis O. Burzio

Published in: Cellular Oncology | Issue 4/2011

Login to get access

Abstract

Background

We have previously shown a differential expression of a family of mitochondrial ncRNAs in normal and cancer cells. Normal proliferating cells and cancer cells express the sense mitochondrial ncRNA (SncmtRNA). In addition, while normal proliferating cells express two antisense mitochondrial ncRNAs (ASncmtRNAs-1 and −2), these transcripts seem to be universally down-regulated in cancer cells. In situ hybridization (ISH) of some normal and cancer tissues reveals nuclear localization of these transcripts suggesting that they are exported from mitochondria.

Methods

FISH and confocal microscopy, in situ digestion with RNase previous to ISH and electron microscopy ISH was employed to confirm the extra-mitochondrial localization of the SncmtRNA and the ASncmtRNAs in normal proliferating and cancer cells of human and mouse.

Results

In normal human kidney and mouse testis the SncmtRNA and the ASncmtRNAs were found outside the organelle and especially localized in the nucleus associated to heterochromatin. In cancer cells, only the SncmtRNA was expressed and was found associated to heterochromatin and nucleoli.

Conclusion

The ubiquitous localization of these mitochondrial transcripts in the nucleus suggests that they are new players in the mitochondrial-nuclear communication pathway or retrograde signaling. Down regulation of the ASncmtRNAs seems to be an important step on neoplastic transformation and cancer progression.
Literature
1.
go back to reference O. Warburg, On respiratory impairment in cancer cells. Science 124, 269–270 (1956)PubMed O. Warburg, On respiratory impairment in cancer cells. Science 124, 269–270 (1956)PubMed
2.
go back to reference J.S. Carew, P. Huang, Mitochondrial defects in cancer. Mol Cancer 1, 1–9 (2002)CrossRef J.S. Carew, P. Huang, Mitochondrial defects in cancer. Mol Cancer 1, 1–9 (2002)CrossRef
3.
go back to reference D.C. Wallace, W.W. Fan, The pathophysiology of mitochondrial disease as modeled in the mouse. Genes Dev 23, 1714–1736 (2009)PubMedCrossRef D.C. Wallace, W.W. Fan, The pathophysiology of mitochondrial disease as modeled in the mouse. Genes Dev 23, 1714–1736 (2009)PubMedCrossRef
4.
go back to reference G. Biswas, O.A. Adebanjo, B.D. Freedman, H.K. Anandatheerthavarada, C. Vijayasarathy, M. Zaidi, M. Kotlikoff, N.G. Avadhani, Retrograde Ca2+ signaling in C2C12 skeletal myocytes in response to mitochondrial genetic and metabolic stress: a novel mode of inter-organelle crosstalk. EMBO J 18, 522–533 (1999)PubMedCrossRef G. Biswas, O.A. Adebanjo, B.D. Freedman, H.K. Anandatheerthavarada, C. Vijayasarathy, M. Zaidi, M. Kotlikoff, N.G. Avadhani, Retrograde Ca2+ signaling in C2C12 skeletal myocytes in response to mitochondrial genetic and metabolic stress: a novel mode of inter-organelle crosstalk. EMBO J 18, 522–533 (1999)PubMedCrossRef
5.
go back to reference G. Biswas, H.K. Anandatheerthavarada, N.G. Avadhani, Mechanism of mitochondrial stress-induced resistance to apoptosis in mitochondrial DNA-depleted C2C12 myocytes. Cell Death Differ 12, 266–278 (2005)PubMedCrossRef G. Biswas, H.K. Anandatheerthavarada, N.G. Avadhani, Mechanism of mitochondrial stress-induced resistance to apoptosis in mitochondrial DNA-depleted C2C12 myocytes. Cell Death Differ 12, 266–278 (2005)PubMedCrossRef
6.
go back to reference G. Amuthan, G. Biswas, S.Y. Zhang, A. Klein-Szanto, C. Vijayasarathy, N.G. Avadhani, Mitochondria-to-nucleus stress signaling induces phenotypic changes, tumor progression and cell invasion. EMBO J 20, 1910–1920 (2001)PubMedCrossRef G. Amuthan, G. Biswas, S.Y. Zhang, A. Klein-Szanto, C. Vijayasarathy, N.G. Avadhani, Mitochondria-to-nucleus stress signaling induces phenotypic changes, tumor progression and cell invasion. EMBO J 20, 1910–1920 (2001)PubMedCrossRef
7.
go back to reference G. Amuthan, G. Biswas, H.K. Ananadatheerthavarada, C. Vijayasarathy, H.M. Shephard, N.G. Avadhani, Mitochondrial stress-induced calcium signaling, phenotypic changes and invasive behavior in human lung carcinoma A549 cells. Oncogene 21, 7839–7849 (2002)PubMedCrossRef G. Amuthan, G. Biswas, H.K. Ananadatheerthavarada, C. Vijayasarathy, H.M. Shephard, N.G. Avadhani, Mitochondrial stress-induced calcium signaling, phenotypic changes and invasive behavior in human lung carcinoma A549 cells. Oncogene 21, 7839–7849 (2002)PubMedCrossRef
8.
go back to reference R.A. Butow, N.G. Avadhani, Mitochondrial signaling: the retrograde response. Mol. Cell 14, 1–15 (2004)PubMedCrossRef R.A. Butow, N.G. Avadhani, Mitochondrial signaling: the retrograde response. Mol. Cell 14, 1–15 (2004)PubMedCrossRef
9.
10.
go back to reference M.T. Ryan, N.J. Hoogenraad, Mitochondrial-nuclear communications. Ann Rev Biochem 76, 701–722 (2007)PubMedCrossRef M.T. Ryan, N.J. Hoogenraad, Mitochondrial-nuclear communications. Ann Rev Biochem 76, 701–722 (2007)PubMedCrossRef
11.
go back to reference G. Biswas, H.K. Anandatheerthavarada, M. Zaidi, N.G. Avadhani, Mitochondria to nucleus stress signaling: a distinctive mechanism of NFkappaB/Rel activation through calcineurin-mediated inactivation of IkappaBbeta. J Cell Biol 161, 507–519 (2003)PubMedCrossRef G. Biswas, H.K. Anandatheerthavarada, M. Zaidi, N.G. Avadhani, Mitochondria to nucleus stress signaling: a distinctive mechanism of NFkappaB/Rel activation through calcineurin-mediated inactivation of IkappaBbeta. J Cell Biol 161, 507–519 (2003)PubMedCrossRef
12.
go back to reference M. Guha, S. Srinivasan, G. Biswas, N.G. Avadhani, Activation of a novel calcineurin-mediated insulin-like growth factor-1 receptor pathway, altered metabolism, and tumor cell invasion in cells subjected to mitochondrial respiratory stress. J Biol Chem 282, 14536–14546 (2007)PubMedCrossRef M. Guha, S. Srinivasan, G. Biswas, N.G. Avadhani, Activation of a novel calcineurin-mediated insulin-like growth factor-1 receptor pathway, altered metabolism, and tumor cell invasion in cells subjected to mitochondrial respiratory stress. J Biol Chem 282, 14536–14546 (2007)PubMedCrossRef
13.
go back to reference S.M. Schieke, T. Finkel, Mitochondrial signaling, TOR and life span. Biol Chem 387, 1357–1361 (2006)PubMedCrossRef S.M. Schieke, T. Finkel, Mitochondrial signaling, TOR and life span. Biol Chem 387, 1357–1361 (2006)PubMedCrossRef
14.
go back to reference G. Biswas, S. Srinivasan, H.K. Anandatheerthavarada, N.G. Avadhani, Dioxin-mediated tumor progression through activation of mitochondria-to-nucleus stress signaling. Proc Natl Acad Sci U S A 105, 186–191 (2008)PubMedCrossRef G. Biswas, S. Srinivasan, H.K. Anandatheerthavarada, N.G. Avadhani, Dioxin-mediated tumor progression through activation of mitochondria-to-nucleus stress signaling. Proc Natl Acad Sci U S A 105, 186–191 (2008)PubMedCrossRef
15.
go back to reference J. Villegas, V.A. Burzio, C. Villota, E. Landerer, R. Martinez, R. Pinto, M.I. Vera, J. Castillo, L.O. Burzio, Expression of a novel non-coding mitochondrial RNA in human proliferating cells. Nucleic Acids Res 35, 7336–7347 (2007)PubMedCrossRef J. Villegas, V.A. Burzio, C. Villota, E. Landerer, R. Martinez, R. Pinto, M.I. Vera, J. Castillo, L.O. Burzio, Expression of a novel non-coding mitochondrial RNA in human proliferating cells. Nucleic Acids Res 35, 7336–7347 (2007)PubMedCrossRef
16.
go back to reference V. Burzio, C. Villota, J. Villegas, E. Landerer, E. Boccardo, L.L. Villa, R. Martinez, C. Lopez, F. Gaete, V. Toro, X. Rodriguez, L.O. Burzio, Expression of a family of noncoding mitocondrial RNAs distinguishes normal from cancer cells. Proc Natl Acad Sci, USA 106, 9430–9434 (2009)PubMedCrossRef V. Burzio, C. Villota, J. Villegas, E. Landerer, E. Boccardo, L.L. Villa, R. Martinez, C. Lopez, F. Gaete, V. Toro, X. Rodriguez, L.O. Burzio, Expression of a family of noncoding mitocondrial RNAs distinguishes normal from cancer cells. Proc Natl Acad Sci, USA 106, 9430–9434 (2009)PubMedCrossRef
17.
go back to reference N.D. Bonawitz, D.A. Clayton, G.S. Shadel, Initiation and beyond: multiple functions of the human mitochondrial transcription machinery. Mol Cell 24, 813–825 (2006)PubMedCrossRef N.D. Bonawitz, D.A. Clayton, G.S. Shadel, Initiation and beyond: multiple functions of the human mitochondrial transcription machinery. Mol Cell 24, 813–825 (2006)PubMedCrossRef
18.
go back to reference M. Falkenberg, N.G. Larsson, C.M. Gustafsson, DNA replication and transcription in mammalian mitochondria. Ann Rev Biochem 76(30), 1–30 (2007) M. Falkenberg, N.G. Larsson, C.M. Gustafsson, DNA replication and transcription in mammalian mitochondria. Ann Rev Biochem 76(30), 1–30 (2007)
19.
go back to reference B. Goic, J. Bustamante, A. Miquel, M. Alvarez, M.I. Vera, P.D. Valenzuela, L.O. Burzio, The nucleoprotein and the viral RNA of infectious salmon anemia virus (ISAV) are localized in the nucleolus of infected cells. Virology 379, 55–63 (2008)PubMedCrossRef B. Goic, J. Bustamante, A. Miquel, M. Alvarez, M.I. Vera, P.D. Valenzuela, L.O. Burzio, The nucleoprotein and the viral RNA of infectious salmon anemia virus (ISAV) are localized in the nucleolus of infected cells. Virology 379, 55–63 (2008)PubMedCrossRef
20.
go back to reference S. Kobayashi, R. Amikura, M. Okada, Presence of mitochondrial large ribosomal RNA outside mitochondria in germ plasm of Drosophila melanogaster. Science 260, 1521–1524 (1993)PubMedCrossRef S. Kobayashi, R. Amikura, M. Okada, Presence of mitochondrial large ribosomal RNA outside mitochondria in germ plasm of Drosophila melanogaster. Science 260, 1521–1524 (1993)PubMedCrossRef
21.
go back to reference F.J. Iborra, D.A. Jackson, P.R. Cook, The path of RNA through nuclear pores: apparent entry from the sides into specialized pores. J Cell Sci 113, 291–302 (2000)PubMed F.J. Iborra, D.A. Jackson, P.R. Cook, The path of RNA through nuclear pores: apparent entry from the sides into specialized pores. J Cell Sci 113, 291–302 (2000)PubMed
22.
go back to reference D. Cmarko, S.O. Bøe, C. Scassellati, A.M. Szilvay, S. Davanger, X.D. Fu, G. Haukenes, K.H. Kalland, S. Fakan, Rev inhibition strongly affects intracellular distribution of human immunodeficiency virus type 1 RNAs. J Virol 76(2002), 10473–10484 (2002)PubMedCrossRef D. Cmarko, S.O. Bøe, C. Scassellati, A.M. Szilvay, S. Davanger, X.D. Fu, G. Haukenes, K.H. Kalland, S. Fakan, Rev inhibition strongly affects intracellular distribution of human immunodeficiency virus type 1 RNAs. J Virol 76(2002), 10473–10484 (2002)PubMedCrossRef
23.
go back to reference R. Gelfand, G. Attardi, Synthesis and turnover of mitochondrial ribonucleic acid in HeLa cells: the mature ribosomal and messenger ribonucleic acid species are metabolically unstable. Mol Cell Biol 1, 497–511 (1981)PubMed R. Gelfand, G. Attardi, Synthesis and turnover of mitochondrial ribonucleic acid in HeLa cells: the mature ribosomal and messenger ribonucleic acid species are metabolically unstable. Mol Cell Biol 1, 497–511 (1981)PubMed
24.
go back to reference R.S. Puranam, G. Attardi, The RNase P associated with HeLa cell mitochondria contains an essential RNA component identical in sequence to that of the nuclear RNase P. Mol Cell Biol 21, 548–561 (2001)PubMedCrossRef R.S. Puranam, G. Attardi, The RNase P associated with HeLa cell mitochondria contains an essential RNA component identical in sequence to that of the nuclear RNase P. Mol Cell Biol 21, 548–561 (2001)PubMedCrossRef
25.
go back to reference G. Wang, H.W. Chen, Y. Oktay, J. Zhang, E.L. Allen, G.M. Smith, K.C. Fan, J.S. Hong, S.W. French, J.M. McCaffery, R.N. Lightowlers, H.C. Morse, C.M. Kehler, M.A. Teitell, PNPASE regulates RNA import into mitochondria. Cell 142, 456–467 (2010)PubMedCrossRef G. Wang, H.W. Chen, Y. Oktay, J. Zhang, E.L. Allen, G.M. Smith, K.C. Fan, J.S. Hong, S.W. French, J.M. McCaffery, R.N. Lightowlers, H.C. Morse, C.M. Kehler, M.A. Teitell, PNPASE regulates RNA import into mitochondria. Cell 142, 456–467 (2010)PubMedCrossRef
26.
go back to reference M.P. King, G. Attardi, Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. Science 246, 500–503 (1989)PubMedCrossRef M.P. King, G. Attardi, Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. Science 246, 500–503 (1989)PubMedCrossRef
27.
go back to reference T. Hayakawa, M. Noda, K. Yasuda, H. Yorifuji, S. Taniguchi, I. Miwa, H. Sakura, Y. Terauchi, J. Hayashi, G.W. Sharp, Y. Kanazawa, Y. Akanuma, Y. Yazaki, T. Kadowaki, Ethidium bromide-induced inhibition of mitochondrial gene transcription suppresses glucose-stimulated insulin release in the mouse pancreatic b-cell line bHC9. J Biol Chem 273, 20300–20307 (1998)PubMedCrossRef T. Hayakawa, M. Noda, K. Yasuda, H. Yorifuji, S. Taniguchi, I. Miwa, H. Sakura, Y. Terauchi, J. Hayashi, G.W. Sharp, Y. Kanazawa, Y. Akanuma, Y. Yazaki, T. Kadowaki, Ethidium bromide-induced inhibition of mitochondrial gene transcription suppresses glucose-stimulated insulin release in the mouse pancreatic b-cell line bHC9. J Biol Chem 273, 20300–20307 (1998)PubMedCrossRef
28.
go back to reference J.E. Sligh, S.E. Levy, K.G. Waymire, P. Allard, D.L. Dillehay, S. Nusinowitz, J.R. Heckenlively, G.R. MacGregor, D.C. Wallace, Maternal germ-line transmission of mutant mtDNAs from embryonic stem cell-derived chimeric mice. Proc Natl Acad, Sci USA 97, 14461–14466 (2000)CrossRef J.E. Sligh, S.E. Levy, K.G. Waymire, P. Allard, D.L. Dillehay, S. Nusinowitz, J.R. Heckenlively, G.R. MacGregor, D.C. Wallace, Maternal germ-line transmission of mutant mtDNAs from embryonic stem cell-derived chimeric mice. Proc Natl Acad, Sci USA 97, 14461–14466 (2000)CrossRef
29.
go back to reference Q. Felty, K.P. Singh, D. Roy, Estrogen-induced G1/S transition of G0 arrested estrogen-dependent breast cancer cells is regulated by mitochondrial oxidant signaling. Oncogene 24, 4883–4893 (2005)PubMedCrossRef Q. Felty, K.P. Singh, D. Roy, Estrogen-induced G1/S transition of G0 arrested estrogen-dependent breast cancer cells is regulated by mitochondrial oxidant signaling. Oncogene 24, 4883–4893 (2005)PubMedCrossRef
30.
go back to reference A. Rego, P.B. Sinclair, W. Tao, I. Kireev, A.S. Belmont, The facultative heterochromatin of the inactive X chromosome has a distinctive condensed ultrastructure. J Cell Sci 121, 1119–1127 (2008)PubMedCrossRef A. Rego, P.B. Sinclair, W. Tao, I. Kireev, A.S. Belmont, The facultative heterochromatin of the inactive X chromosome has a distinctive condensed ultrastructure. J Cell Sci 121, 1119–1127 (2008)PubMedCrossRef
31.
go back to reference A. Chacinska, C.M. Koehler, D. Milenkovic, T. Lithgow, N. Pfanner, Importing mitochondrial proteins: machineries and mechanisms. Cell 138, 628–644 (2009)PubMedCrossRef A. Chacinska, C.M. Koehler, D. Milenkovic, T. Lithgow, N. Pfanner, Importing mitochondrial proteins: machineries and mechanisms. Cell 138, 628–644 (2009)PubMedCrossRef
32.
go back to reference D.J. Pagliarini, S.E. Calvo, B. Chang, S.A. Sheth, S.B. Vafai, S.E. Ong, G.A. Walford, C. Sugiana, A. Boneh, W.K. Chen, D.E. Hill, M. Vidal, J.G. Evans, D.R. Thorburn, S.A. Carr, V.K. Mootha, A mitochondrial protein compendium elucidates complex I disease biology. Cell 134, 112–123 (2008)PubMedCrossRef D.J. Pagliarini, S.E. Calvo, B. Chang, S.A. Sheth, S.B. Vafai, S.E. Ong, G.A. Walford, C. Sugiana, A. Boneh, W.K. Chen, D.E. Hill, M. Vidal, J.G. Evans, D.R. Thorburn, S.A. Carr, V.K. Mootha, A mitochondrial protein compendium elucidates complex I disease biology. Cell 134, 112–123 (2008)PubMedCrossRef
33.
go back to reference J.D. Alfonzo, D. Söll, Mitochondrial tRNA import–the challenge to understand has just begun. Biol Chem 390, 717–722 (2009)PubMedCrossRef J.D. Alfonzo, D. Söll, Mitochondrial tRNA import–the challenge to understand has just begun. Biol Chem 390, 717–722 (2009)PubMedCrossRef
34.
go back to reference A. Smirnov, C. Comte, A.-M. Mager-Heckel, V. Addis, I.A. Krasheninnikov, R.P. Martin, E. Entelis, I. Tarassov, Mitochondrial enzyme rhodanese is essential for 5 S ribosomal RNA import into human mitochondria. J Biol Chem 285(2010), 30792–30803 (2010)PubMedCrossRef A. Smirnov, C. Comte, A.-M. Mager-Heckel, V. Addis, I.A. Krasheninnikov, R.P. Martin, E. Entelis, I. Tarassov, Mitochondrial enzyme rhodanese is essential for 5 S ribosomal RNA import into human mitochondria. J Biol Chem 285(2010), 30792–30803 (2010)PubMedCrossRef
35.
go back to reference B.T. Kren, P.Y. Wong, A. Sarver, X. Zhang, Y. Zeng, C.J. Steer, MicroRNAs identified in highly purified liver-derived mitochondria may play a role in apoptosis. RNA Biol 6, 65–72 (2009)PubMedCrossRef B.T. Kren, P.Y. Wong, A. Sarver, X. Zhang, Y. Zeng, C.J. Steer, MicroRNAs identified in highly purified liver-derived mitochondria may play a role in apoptosis. RNA Biol 6, 65–72 (2009)PubMedCrossRef
Metadata
Title
Nuclear localization of the mitochondrial ncRNAs in normal and cancer cells
Authors
Eduardo Landerer
Jaime Villegas
Veronica A. Burzio
Luciana Oliveira
Claudio Villota
Constanza Lopez
Franko Restovic
Ronny Martinez
Octavio Castillo
Luis O. Burzio
Publication date
01-08-2011
Publisher
Springer Netherlands
Published in
Cellular Oncology / Issue 4/2011
Print ISSN: 2211-3428
Electronic ISSN: 2211-3436
DOI
https://doi.org/10.1007/s13402-011-0018-8

Other articles of this Issue 4/2011

Cellular Oncology 4/2011 Go to the issue
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine