title: eEF1Bγ binds the Che-1 and TP53 gene promoters and their transcripts.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: CNR-Institute of Molecular Biology and Pathology, Department of Molecular Medicine, Sapienza University, Viale Regina Elena 291, 00161, Rome, Italy. cinzia.pisani@uniroma1.it.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Pisani
authors.affiliation: CNR-Institute of Molecular Biology and Pathology, Department of Molecular Medicine, Sapienza University, Viale Regina Elena 291, 00161, Rome, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Onori
authors.affiliation: CNR -Institute of Cell Biology and Neurobiology, Rome, Italy.|IRCCS Fondazione Santa Lucia, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Gabanella
authors.affiliation: CNR-Institute of Molecular Biology and Pathology, Department of Molecular Medicine, Sapienza University, Viale Regina Elena 291, 00161, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Delle Monache
authors.affiliation: CNR -Institute of Cell Biology and Neurobiology, Rome, Italy.|IRCCS Fondazione Santa Lucia, Rome, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Borreca
authors.affiliation: CNR -Institute of Cell Biology and Neurobiology, Rome, Italy.|IRCCS Fondazione Santa Lucia, Rome, Italy.
authors.forename: Martine
authors.initials: M
authors.lastname: Ammassari-Teule
authors.affiliation: Department of Research, Advanced Diagnostic, and Technological Innovation, SAFU Laboratory, Regina Elena Cancer Institute, Rome, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Fanciulli
authors.affiliation: CNR -Institute of Cell Biology and Neurobiology, Rome, Italy.|IRCCS Fondazione Santa Lucia, Rome, Italy.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Di Certo
authors.affiliation: CNR-Institute of Molecular Biology and Pathology, Department of Molecular Medicine, Sapienza University, Viale Regina Elena 291, 00161, Rome, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Passananti
authors.affiliation: CNR-Institute of Molecular Biology and Pathology, Department of Molecular Medicine, Sapienza University, Viale Regina Elena 291, 00161, Rome, Italy. nicoletta.corbi@uniroma1.it.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Corbi
mesh_terms: D020413:3' Untranslated Regions; D051017:Apoptosis Regulatory Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000235:genetics; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism*; D020648:Peptide Elongation Factor 1 / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; C402637:AATF protein, human; D051017:Apoptosis Regulatory Proteins; D020648:Peptide Elongation Factor 1; D012333:RNA, Messenger; D012097:Repressor Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C000599475:eEF1B-gamma protein, human
keywords: AATF; Che-1; DNA damage; Mitochondria; POLR2C; RIP assay; RNA binding protein; Translation elongation factor; eEF1Bγ; p53