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Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2016

Open Access 01-12-2016 | Research

eEF1Bγ binds the Che-1 and TP53 gene promoters and their transcripts

Authors: Cinzia Pisani, Annalisa Onori, Francesca Gabanella, Francesca Delle Monache, Antonella Borreca, Martine Ammassari-Teule, Maurizio Fanciulli, Maria Grazia Di Certo, Claudio Passananti, Nicoletta Corbi

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2016

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Abstract

Background

We have previously shown that the eukaryotic elongation factor subunit 1B gamma (eEF1Bγ) interacts with the RNA polymerase II (pol II) alpha-like subunit “C” (POLR2C), alone or complexed, in the pol II enzyme. Moreover, we demonstrated that eEF1Bγ binds the promoter region and the 3’ UTR mRNA of the vimentin gene. These events contribute to localize the vimentin transcript and consequentially its translation, promoting a proper mitochondrial network.

Methods

With the intent of identifying additional transcripts that complex with the eEF1Bγ protein, we performed a series of ribonucleoprotein immunoprecipitation (RIP) assays using a mitochondria-enriched heavy membrane (HM) fraction.

Results

Among the eEF1Bγ complexed transcripts, we found the mRNA encoding the Che-1/AATF multifunctional protein. As reported by other research groups, we found the tumor suppressor p53 transcript complexed with the eEF1Bγ protein. Here, we show for the first time that eEF1Bγ binds not only Che-1 and p53 transcripts but also their promoters. Remarkably, we demonstrate that both the Che-1 transcript and its translated product localize also to the mitochondria and that eEF1Bγ depletion strongly perturbs the mitochondrial network and the correct localization of Che-1. In a doxorubicin (Dox)-induced DNA damage assay we show that eEF1Bγ depletion significantly decreases p53 protein accumulation and slightly impacts on Che-1 accumulation. Importantly, Che-1 and p53 proteins are components of the DNA damage response machinery that maintains genome integrity and prevents tumorigenesis.

Conclusions

Our data support the notion that eEF1Bγ, besides its canonical role in translation, is an RNA-binding protein and a key player in cellular stress responses. We suggest for eEF1Bγ a role as primordial transcription/translation factor that links fundamental steps from transcription control to local translation.
Appendix
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Literature
1.
Merrick WC, Nyborg J. The protein biosynthesis elongation cycle. In Translational Control of Gene Expression. CSHL Press NY (Sonenberg N, Hershey J, Mathews M (Eds)). 2000; 89–125.
2.
Le Sourd F, Boulben S, Le Bouffant R, Cormier P, Morales J, Belle R, Mulner-Lorillon O. eEF1B: At the dawn of the 21st century. Biochim Biophys Acta. 2006;1759(1–2):13–31.CrossRefPubMed
3.
Trosiuk TV, Shalak VF, Szczepanowski RH, Negrutskii BS, El'skaya AV. A non-catalytic N-terminal domain negatively influences the nucleotide exchange activity of translation elongation factor 1Bα. FEBS J. 2016;283(3):484–97.CrossRefPubMed
4.
Esposito AM, Kinzy TG. The eukaryotic translation elongation Factor 1Bgamma has a non-guanine nucleotide exchange factor role in protein metabolism. J Biol Chem. 2010;285(49):37995–8004.CrossRefPubMedPubMedCentral
5.
Corbi N, Batassa EM, Pisani C, Onori A, Di Certo MG, Strimpakos G, Fanciulli M, Mattei E, Passananti C. The eEF1γ subunit contacts RNA polymerase II and binds vimentin promoter region. PLoS One. 2010;5(12):e14481.CrossRefPubMedPubMedCentral
6.
7.
Veremieva M, Kapustian L, Khoruzhenko A, Zakharychev V, Negrutskii B, El’skaya A. Independent overexpression of the subunits of translation elongation factor complex eEF1H in human lung cancer. BMC Cancer. 2014;14:913.CrossRefPubMedPubMedCentral
8.
Al-Maghrebi M, Anim JT, Olalu AA. Up-regulation of eukaryotic elongation factor-1 subunits in breast carcinoma. Anticancer Res. 2005;25(3c):2573–7.PubMed
9.
Chi K, Jones DV, Frazier ML. Expression of an elongation factor 1 gamma-related sequence in adenocarcinomas of the colon. Gastroenterology. 1992;103(1):98–102.CrossRefPubMed
10.
Mimori K, Mori M, Tanaka S, Akiyoshi T, Sugimachi K. The overexpression of elongation factor 1 gamma mRNA in gastric carcinoma. Cancer. 1995;75(6 Suppl):1446–9.CrossRefPubMed
11.
Lew Y, Jones DV, Mars WM, Evans D, Byrd D, Frazier ML. Expression of elongation factor-1 gamma-related sequence in human pancreatic cancer. Pancreas. 1992;7(2):144–52.CrossRefPubMed
12.
Ruggero D. Translational control in cancer etiology. Cold Spring Harb Perspect Biol. 2013;5(2).
13.
Chen B, Tan Z, Gao J, Wu W, Liu L, Jin W, Cao Y, Zhao S, Zhang W, Qiu Z, Liu D, Mo X, Li W. Hyperphosphorylation of ribosomal protein S6 predicts unfavorable clinical survival in non-small cell lung cancer. J Exp Clin Cancer Res. 2015;34:126.CrossRefPubMedPubMedCentral
14.
15.
16.
Mimori K, Mori M, Inoue H, Ueo H, Mafune K, Akiyoshi T, Sugimachi K. Elongation factor 1 gamma mRNA expression in oesophageal carcinoma. Gut. 1996;38(1):66–70.CrossRefPubMedPubMedCentral
17.
Kim S, Kellner J, Lee CH, Coulombe PA. Interaction between the keratin cytoskeleton and eEF1Bgamma affects protein synthesis in epithelial cells. Nat Struct Mol Biol. 2007;14(10):982–3.CrossRefPubMed
18.
Al-Maghrebi M, Brulé H, Padkina M, Allen C, Holmes WM, Zehner ZE. The 3’ untranslated region of human vimentin mRNA interacts with protein complexes containing eEF-1gamma and HAX-1. Nucleic Acids Res. 2002;30(23):5017–28.CrossRefPubMedPubMedCentral
19.
Shi Y, Di Giammartino DC, Taylor D, Sarkeshik A, Rice WJ, Yates 3rd JR, Frank J, Manley JL. Molecular architecture of the human pre-mRNA 3’ processing complex. Mol Cell. 2009;33(3):365–76.CrossRefPubMedPubMedCentral
20.
Tang HL, Lung HL, Wu KC, Le AH, Tang HM, Fung MC. Vimentin supports mitochondrial morphology and organization. Biochem J. 2008;410(1):141–6.CrossRefPubMed
21.
Chernoivanenko IS, Matveeva EA, Gelfand VI, Goldman RD, Minin AA. Mitochondrial membrane potential is regulated by vimentin intermediate filaments. FASEB J. 2015;29(3):820–7.CrossRefPubMed
22.
Jo JH, Chung TM, Youn H, Yoo JY. Cytoplasmic parafibromin/hCdc73 targets and destabilizes p53 mRNA to control p53-mediated apoptosis. Nat Commun. 2014;5:5433.CrossRefPubMed
23.
Liu D, Sheng C, Gao S, Jiang W, Li J, Yao C, Chen H, Wu J, Chen S, Huang W. eEF1Bγ is a positive regulator of NF-кB signaling pathway. Biochem Biophys Res Commun. 2014;446(2):523–8.CrossRefPubMed
24.
Matassa DS, Amoroso MR, Agliarulo I, Maddalena F, Sisinni L, Paladino S, Romano S, Romano MF, Sagar V, Loreni F, Landriscina M, Esposito F. Translational control in the stress adaptive response of cancer cells: a novel role for the heat shock protein TRAP1. Cell Death Dis. 2013;4:e851.CrossRefPubMedPubMedCentral
25.
Sainsbury S, Bernecky C, Cramer P. Structural basis of transcription initiation by RNA polymerase II. Nat Rev Mol Cell Biol. 2015;16(3):129–43.CrossRefPubMed
26.
Kornberg RD. Mediator and the mechanism of transcriptional activation. Trends Biochem Sci. 2005;30(5):235–9.CrossRefPubMed
27.
Fanciulli M, Bruno T, Di Padova M, De Angelis R, Lovari S, Floridi A, Passananti C. The interacting RNA polymerase II subunits, hRPB11 and hRPB3, are coordinately expressed in adult human tissues and down-regulated by doxorubicin. FEBS Lett. 1998;427(2):236–40.CrossRefPubMed
28.
Davis JA, Takagi Y, Kornberg RD, Asturias FA. Structure of the yeast RNA polymerase II holoenzyme: Mediator conformation and polymerase interaction. Mol Cell. 2002;10(2):409–15.CrossRefPubMed
29.
Woychik NA, McKune K, Lane WS, Young RA. Yeast RNA polymerase II subunit RPB11 is related to a subunit shared by RNA polymerase I and III. Gene Expr. 1993;3(1):77–82.PubMed
30.
Corbi N, Di Padova M, De Angelis R, Bruno T, Libri V, Iezzi S, Floridi A, Fanciulli M, Passananti C. The alpha-like RNA polymerase II core subunit 3 (RPB3) is involved in tissue-specific transcription and muscle differentiation via interaction with the myogenic factor myogenin. FASEB J. 2002;16(12):1639–41.PubMed
31.
Harden TT, Wells CD, Friedman LJ, Landick R, Hochschild A, Kondev J, Gelles J. Bacterial RNA polymerase can retain σ70 throughout transcription. Proc Natl Acad Sci U S A. 2016;113(3):602–7.CrossRefPubMedPubMedCentral
32.
Passananti C, Fanciulli M. The anti-apoptotic factor Che-1/AATF links transcriptional regulation, cell cycle control, and DNA damage response. Cell Div. 2007;2:21.CrossRefPubMedPubMedCentral
33.
34.
35.
36.
37.
Desantis A, Bruno T, Catena V, De Nicola F, Goeman F, Iezzi S, Sorino C, Gentileschi MP, Germoni S, Monteleone V, Pellegrino M, Kann M, De Meo PD, Pallocca M, Höpker K, Moretti F, Mattei E, Reinhardt HC, Floridi A, Passananti C, Benzing T, Blandino G, Fanciulli M. Che-1 modulates the decision between cell cycle arrest and apoptosis by its binding to p53. Cell Death Dis. 2015;6:e1764.CrossRefPubMedPubMedCentral
38.
Bruno T, Desantis A, Bossi G, Di Agostino S, Sorino C, De Nicola F, Iezzi S, Franchitto A, Benassi B, Galanti S, La Rosa F, Floridi A, Bellacosa A, Passananti C, Blandino G, Fanciulli M. Che-1 promotes tumor cell survival by sustaining mutant p53 transcription and inhibiting DNA damage response activation. Cancer Cell. 2010;18(2):122–34.CrossRefPubMed
39.
Karbowski M, Neutzner A, Youle RJ. The mitochondrial E3 ubiquitin ligase MARCH5 is required for Drp1 dependent mitochondrial division. J Cell Biol. 2007;178(1):71–84.CrossRefPubMedPubMedCentral
40.
Keene JD, Komisarow JM, Friedersdorf MB. RIP-Chip: the isolation and identification of mRNAs, microRNAs and protein components of ribonucleoprotein complexes from cell extracts. Nat Protoc. 2006;1(1):302–7.CrossRefPubMed
41.
Onori A, Pisani C, Strimpakos G, Monaco L, Mattei E, Passananti C, Corbi N. UtroUp is a novel six zinc finger artificial transcription factor that recognises 18 base pairs of the utrophin promoter and efficiently drives utrophin upregulation. BMC Mol Biol. 2013;14:3.CrossRefPubMedPubMedCentral
42.
Gabanella F, Pisani C, Borreca A, Farioli-Vecchioli S, Ciotti MT, Ingegnere T, Onori A, Ammassari-Teule M, Corbi N, Canu N, Monaco L, Passananti C, Di Certo MG. SMN affects membrane remodelling and anchoring of the protein synthesis machinery. J Cell Sci. 2016;129(4):804–16.CrossRefPubMed
43.
Raj A, Tyagi S. Detection of individual endogenous RNA transcripts in situ using multiple singly labeled probes. Methods Enzymol. 2010;472:365–86.CrossRefPubMed
44.
45.
Li Q, Lau A, Morris TJ, Guo L, Fordyce CB, Stanley EF. A syntaxin 1, Galpha(o), and N-type calcium channel complex at a presynaptic nerve terminal: analysis by quantitative immunocolocalization. J Neurosci. 2004;24(16):4070–81.CrossRefPubMed
46.
Fanciulli M, Bruno T, Di Padova M, De Angelis R, Iezzi S, Iacobini C, Floridi A, Passananti C. Identification of a novel partner of RNA polymerase II subunit 11, Che-1, which interacts with and affects the growth suppression function of Rb. FASEB J. 2000;14(7):904–12.PubMed
47.
Zehner ZE, Shepherd RK, Gabryszuk J, Fu TF, Al-Ali M, Holmes WM. RNA-protein interactions within the 3 ‘untranslated region of vimentin mRNA. Nucleic Acids Res. 1997;25(16):3362–70.CrossRefPubMedPubMedCentral
48.
Bermano G, Shepherd RK, Zehner ZE, Hesketh JE. Perinuclear mRNA localisation by vimentin 3'-untranslated region requires a 100 nucleotide sequence and intermediate filaments. FEBS Lett. 2001;497(2–3):77–81.CrossRefPubMed
49.
Sasvari Z, Izotova L, Kinzy TG, Nagy PD. Synergistic roles of eukaryotic translation elongation factors 1Bγ and 1A in stimulation of tombusvirus minus-strand synthesis. PLoS Pathog. 2011;7(12):e1002438.CrossRefPubMedPubMedCentral
50.
51.
Querido E, Chartrand P. Using fluorescent proteins to study mRNA trafficking in living cells. Methods Cell Biol. 2008;85:273–92.CrossRefPubMed
52.
Sanchez G, Dury AY, Murray LM, Biondi O, Tadesse H, El Fatimy R, Kothary R, Charbonnier F, Khandjian EW, Côté J. A novel function for the survival motoneuron protein as a translational regulator. Hum Mol Genet. 2013;22(4):668–84.CrossRefPubMed
53.
Dahl JA, Collas P. A rapid micro chromatin immunoprecipitation assay (microChIP). Nat Protoc. 2008;3(6):1032–45.CrossRefPubMed
54.
Sorino C, Bruno T, Desantis A, Di Certo MG, Iezzi S, De Nicola F, Catena V, Floridi A, Chessa L, Passananti C, Cundari E, Fanciulli M. Centrosomal Che-1 protein is involved in the regulation of mitosis and DNA damage response by mediating pericentrin (PCNT)-dependent Chk1 protein localization. J Biol Chem. 2013;288(32):23348–57.CrossRefPubMedPubMedCentral
55.
Ishigaki S, Fonseca SG, Oslowski CM, Jurczyk A, Shearstone JR, Zhu LJ, Permutt MA, Greiner DL, Bortell R, Urano F. AATF mediates an antiapoptotic effect of the unfolded protein response through transcriptional regulation of AKT1. Cell Death Differ. 2010;17(5):774–86.CrossRefPubMed
56.
Williams AB, Schumacher B. p53 in the DNA-Damage-Repair Process. Cold Spring Harb Perspect Med. 2016;6(5).
57.
Xie J, Guo Q. Apoptosis antagonizing transcription factor protects renal tubule cells against oxidative damage and apoptosis induced by ischemia-reperfusion. J Am Soc Nephrol. 2006;17(12):3336–46.CrossRefPubMed
58.
59.
Kaul D, Sharma S, Garg A. Mitochondrial uncoupling protein (UCP2) gene expression is regulated by miR-2909. Blood Cells Mol Dis. 2015;55(1):89–93.CrossRefPubMed
60.
Kamp WM, Wang PY, Hwang PM. TP53 mutation, mitochondria and cancer. Curr Opin Genet Dev. 2016;38:16–22.CrossRefPubMed
61.
Park JH, Zhuang J, Li J, Hwang PM. p53 as guardian of the mitochondrial genome. FEBS Lett. 2016;590(7):924–34.CrossRefPubMed
62.
Qin LS, Jia PF, Zhang ZQ, Zhang SM. ROS-p53-cyclophilin-D signaling mediates salinomycin-induced glioma cell necrosis. J Exp Clin Cancer Res. 2015;34:57.CrossRefPubMedPubMedCentral
63.
Dutertre M, Lambert S, Carreira A, Amor-Guéret M, Vagner S. DNA damage: RNA-binding proteins protect from near and far. Trends Biochem Sci. 2014;39(3):141–9.CrossRefPubMed
64.
Truitt ML, Ruggero D. New frontiers in translational control of the cancer genome. Nat Rev Cancer. 2016;16(5):288–304.CrossRefPubMed
65.
Metadata
Title
eEF1Bγ binds the Che-1 and TP53 gene promoters and their transcripts
Authors
Cinzia Pisani
Annalisa Onori
Francesca Gabanella
Francesca Delle Monache
Antonella Borreca
Martine Ammassari-Teule
Maurizio Fanciulli
Maria Grazia Di Certo
Claudio Passananti
Nicoletta Corbi
Publication date
01-12-2016
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2016
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/s13046-016-0424-x

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