Atlas of Genetics and Cytogenetics in Oncology and Haematology

OPEN ACCESS JOURNAL INIST-CNRS

Gene Section Review

EEF1G (Eukaryotic elongation factor 1 gamma) Luigi Cristiano Aesthetic and medical biotechnologies research unit, Prestige, Terranuova Bracciolini, Italy; [email protected]

Published in Atlas Database: March 2019 Online updated version : http://AtlasGeneticsOncology.org/Genes/EEF1GID54272ch11q12.html Printable original version : http://documents.irevues.inist.fr/bitstream/handle/2042/70656/03-2019-EEF1GID54272ch11q12.pdf DOI: 10.4267/2042/70656 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2020 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Abstract Keywords EEF1G; Eukaryotic translation elongation factor 1 Eukaryotic translation elongation factor 1 gamma, gamma; Translation; Translation elongation factor; alias eEF1G, is a that plays a main function protein synthesis; cancer; oncogene; cancer marker in the elongation step of translation process but also covers numerous moonlighting roles. Considering its Identity importance in the cell it is found frequently Other names: EF1G, GIG35, PRO1608, EEF1γ, overexpressed in human cancer cells and thus this EEF1Bγ review wants to collect the state of the art about EEF1G, with insights on DNA, RNA, protein HGNC (Hugo): EEF1G encoded and the diseases where it is implicated. Location: 11q12.3

Figure. 1. Splice variants of EEF1G. The figure shows the locus on 11 of the EEF1G and its splicing variants (grey/blue box). The primary transcript is EEF1G-001 mRNA (green/red box), but also EEF1G-201 variant is able to codify for a protein (reworked from https://www.ncbi.nlm.nih.gov/gene/1937; http://grch37.ensembl.org; www.genecards.org)

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Lenght Lenght Variant Name RefSeq (1) Transcript ID Exons Type RefSeq (2) (bp) (aa) 1 EEF1G-001 NM_001404.5 - 10 Protein coding 1446 NP_001395.1 437 EEF1G-002 (EEF1G- 2 AK092787.1 ENST00000525340.5 9 Retained intron 2496 - - 203) EEF1G-003 (EEF1G- Processed 3 - ENST00000532986.1 5 578 - - 204) transcript EEF1G-004 (EEF1G- Processed 4 - ENST00000524420.5 5 557 - - 202) transcript 5 EEF1G-201 AK300203.1 - 10 Protein coding 1631 BAG61974.1 487

Table.1. Splice variants of EEF1G gene (reworked from http://grch37.ensembl.org)

expressed ubiquitously in human tissues with a DNA/RNA different expression level in relation to the specific tissue type. Minor expression levels are reported for Description brain, liver, lung, pancreas, salivary glands and testis EEF1G (Eukaryotic Translation Elongation Factor 1 while on the contrary a significantly higher Gamma) is a protein coding gene that starts at expression level is found in the ovary (Fagerberg et 62,559,601 nt and ends at 62,573,988 nt from qter al., 2014). Five splice variants for EEF1G were and with a length of 14388 bp. The current reference observed (Table.1): the main reference variant is sequence is NC_000011.10 and contains 10 exons. It EEF1G-001 and the others are EEF1G-002, EEF1G- is proximal to the nucleotidyl transferase TUT1 003, EEF1G-004 and EEF1G-201 (Figure.1). Only (terminal uridylyl transferase 1) gene and to the two of which codify for a protein, i.e. EEF1G-001 AHNAK nucleoprotein gene. Inside the nucleotidic and EEF1G-201, while the others are non-coding sequence of EEF1G there is also a short non-coding RNA sequences (ncRNAs), classified as processed sequence of the microRNA MIR6747 that starts transcripts, i.e. nucleotide sequences that do not from 62567011 bp and ends at 62567071 bp and is contain an open reading frame (ORF) and 61 bp long. Around the genomic locus of EEF1G alternatively spliced transcripts, i.e. retained intron take place different or enhancer sequences. Furthermore, there is a potential transcriptional elements. Two strong elements are readthrough with the inclusion of TUT1 gene. closer to the sequence of EEF1G gene and are located at -1.4 kb and at +2.4 kb respectively and Pseudogene have a high influence of different kind of in According to Gene, the analysis of the human their proximity, such as EEF1G1 and TUT1. genome revealed the presence of nine pseudogenes for EEF1G (Table.2) classified as processed Transcription pseudogenes and probably originated by EEF1G mRNA is 1446 bp long with a reference retrotransposition. If these elements have any sequence reported in GeneBank as NM_001404.5. regulatory roles on the expression of the respective The 5'UTR sequence is not very long and counts 49 gene as described for others (Hirotsune et al., 2003), nt. The CDS is extended from 50 to 1363 nt, while is only speculation in the absence of experimental the 3'UTR starts from 1364 until 1446 nt. EEF1G is evidence.

Gene Gene name Gene ID RefSeq Locus Location Start End Lenght (nt) EEF1GP1 EEF1G1 Pseudogene 1 646837 NC_000007.14 Chromosome 7 7q31.33 125033433 125035389 1957 EEF1GP2 EEF1G1 Pseudogene 2 100130260 NC_000005.10 5q32 147922182 147923422 1241 EEF1GP3 EEF1G1 Pseudogene 3 651628 NC_000003.12 Chromosome 3 3p22.1 40596122 40597519 1398 EEF1GP4 EEF1G1 Pseudogene 4 100129403 NC_000003.12 Chromosome 3 3q26.1 161324837 161326238 1402 EEF1GP5 EEF1G1 Pseudogene 5 642357 NC_000023.11 Chromosome X Xq23 115702791 115704195 1405 EEF1GP6 EEF1G1 Pseudogene 6 100421733 NC_000006.12 Chromosome 6 6q16.1 96750800 96751728 929 EEF1GP7 EEF1G1 Pseudogene 7 645311 NC_000001.11 Chromosome 1 1p32.3 52573115 52573818 704 EEF1GP8 EEF1G1 Pseudogene 8 391698 NC_000004.12 Chromosome 4 4q28.2 129903001 129904244 1244 LOC729998 EEF1G1 Pseudogene 9 729998 NC_000007.14 Chromosome 7 7q33 133034515 133035940 1426

Table.2 EEF1G pseudogene (reworked from https://www.ncbi.nlm.nih.gov/gene/1937)

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EEF1G (Eukaryotic translation elongation factor 1 gamma) Cristiano L

Figure.2 eEF1G protein structure analysis. (1) Primary structure of eEF1G with emphasis on its three principal domains (reworked from Achilonu et al.,2014; https://www.uniprot.org; https://www.proteomicsdb.org; https://prosite.expasy.org); (2) Secondary structure (from https://www.ebi.ac.uk); (3) Tertiary structure: above, front view and below, top view (from http://www.cathdb.info)

however eEF1G seems to be necessary for this Protein nucleotide exchange. Studies did not confirm its Are described two isoforms for eEF1G and it is direct involvement in this process but it is supposed shown that this protein has a glutathione S- that it may stimulate the activity of eEF1B and transferase activity. In addition, eEF1G is a guarantee stability to entire eEF1H complex (Ejiri, structural constituent of a greater protein complex 2002; Mansilla et al., 2002). In addition, studies that is in relation to the and plays a role in revealed that eEF1G sequence does not contain any the elongation step of protein synthesis. consensus sequence for ATP or GTP binding (Maessen et al., 1987). Description There are known two isoforms produced by The eukaryotic elongation factor 1-gamma (alias alternative splicing: the isoform 1 (RefSeq eEF1G, eEF1γ, heEF1γ, eEF1Bγ) is a subunit of the NP_001395.1; UniParc, P26641-1), that has been macromolecular eukaryotic elongation factor-1 chosen as the canonical sequence, is formed by 437 complex (alias eEF1, also called eEF1H), a high- residues and has an overall molecular weight of molecular-weight form made up of an aggregation of 50.12 kDa, while the isoform 2 (RefSeq different protein subunits: eEF1A (alias eEF1α), BAG61974.1; UniParc, P26641-2) is 487 amino eEF1B (alias eEF1 β, eEF1Bα, eEF1B2), eEF1G, acids long with 56.15 kDa of molecular weight. The eEF1D (alias eEF1δ, eEF1Bδ) and valyl t-RNA sequence of this isoform differs from the canonical synthetase (valRS). eEF1H protein complex plays sequence for the substitution of the first four amino central roles in peptide elongation during eukaryotic acids (MAAG) by the insertion of other fifty residues protein biosynthesis, in particular for the delivery of (MAERWVAPAVLRRARFASTFFLSPQIYAHKD aminoacyl-tRNAs to ribosome mediated by the GDLRSAFFILSFKRGEFIPFLNW) with the hydrolysis of GTP. In fact, during the translation creation of an alternative sequence (Ota elongation step, the inactive GDP-bound form of et al., 2004). No experimental data or other studies eEF1A (eEF1A-GDP) is converted to its active were performed for this isoform, so its biological GTP-bound form (eEF1A-GTP) by eEF1BGD role is totally unknown. complex-mediated GTP hydrolysis so it acts as a eEF1G shown hydrophobic properties, has a guanine nucleotide exchange factor (GEF), relatively high isoelectric point (pI ≈ 7) (van Damme regenerating eEF1A-GTP for the successive et al., 1991) and the analysis of its primary and elongation cycle. The physiological role of eEF1G in secondary structures revealed some interesting the translation context is still not well defined, characteristics. In fact, it is a multi-domain protein

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EEF1G (Eukaryotic translation elongation factor 1 gamma) Cristiano L

which consists of three main domains: from the (LARS), cysteine-tRNA synthetase (CARS), leucine amino to carboxyl half terminal there are a zipper putative tumor suppressor 1 (LZTS1), enoyl- glutathione S-transferase (GST)-like N-terminus CoA hydratase 1(ECH1), plasminogen receptor domain (NT-eEF1G), a glutathione S-transferase (PLGRKT), small ubiquitin-related modifier 2 (GST)-like C-terminus domain (CT-eEF1G) and a (SUMO2), ATP-binding cassette sub-family C conserved C-terminal domain (CD-eEF1G) member 9 ( ABCC9), tripartite motif containing 55 (Achilonu et al.,2014). NT-eEF1G and CT-eEF1G (TRIM55), E3 ubiquitin-protein ligase (TRIM63), domains show a homology to the theta class of interleukin enhancer binding factor 2 (ILF2), glutathione S-transferases (GSTs) (Koonin et al., vascular cell adhesion protein 1 (VCAM1), 1994; Janssen and Möller, 1988). eukaryotic translation elongation factor 1 delta The NT-eEF1G domain of eEF1G, by using pseudogene 3 (EEF1DP3), RNA-binding protein 6 secondary structure prediction algorithms, seems to (RBM6) (HuRI database - have a predominant α-helix secondary structure http://interactome.baderlab.org/), ATP-dependent (Achilonu et al.,2014) and it was demonstrated that DNA helicase Q5 (RECQL5) and fasciculation and interacts directly with the N-terminal domain of elongation protein zeta 1 ( FEZ1)(Ishii et al., 2001) eEF1B (van Damme et al., 1991) and also with the although the nature of these interactions are poorly N-terminal domain of eEF1D in independent manner understood. (Cao et al., 2014; Mansilla et al., 2002; Janssen et al., Post-translational modifications. Some post- 1994), although different interactional models were translational modifications are observed, such as proposed (Le Sourd et al., 2006; Jiang et al.,2005; phosphorylation, acetylation and S-nitrosylation. Sheu and Traugh, 1999; Minella et al., 1998). It does 1) Phosphorylation: it was demonstrated that not seem to have a direct interaction with other eEF1G is a target of the kinases, in particular the cell eEF1H elements. cycle protein kinase CDK1 /cyclin B (Le Sourd et In addition, the presence of an enzymatically active al., 2006; Mansilla et al., 2002). GST element could be involved in detoxification of There are at least four positions for phosphorylation: oxygen radicals and the over-expression of eEF1G two on threonine residues (T43, T230) and two on in cancer cells could influence their response to serine residues (S286, S406) (Olsen et al., 2010; oxidative stress and their aggressiveness (Koonin et http://hprd.org). al., 1994). It is assumed that these phosphorylations play a The calculated secondary structure of the CT-eEF1G regulatory role, but their exact functional domain of eEF1G shows both α-helix and β-strand significance is poorly understood. secondary structure elements (Achilonu et al.,2014) 2) Acetylation: there are three positions for and does not interact with eEF1B but instead is the acetylation on lysine residues (K132, K147, K434) candidate domain to has transient interactions with (Choudhary et al., 2009; http://hprd.org). other or cell structures. 3) S-nitrosylation: there is one most probable The CD-eEF1G domain of eEF1G seems to be position for S-Nitrosylation on a cysteine residue mostly in α-helix secondary structure with few β- (C194) (Han and Chen, 2008; http://hprd.org). strands (Achilonu et al.,2014) and currently it does Expression not show particular elements or interactions. It is interesting that eEF1G shows two internal EEF1G mRNA is expressed widely as previous repeats of eight amino acids (VFGEXNXS) at reported while the presence of eEF1G protein in positions 35 - 42 and 355 - 362 respectively, that are human tissues shows unexpected differences i.e. located in its amino-terminal and carboxy-terminal higher levels of protein are observed in cerebellum, halves. hippocampus, esophagus, stomach, small intestine The roles of these two octapeptides are still unknown and pancreas while its low expression levels are even if they could cover the function of binding-sites reported in oral mucosa, bronchus, lung, parathyroid (van Damme et al., 1991; Maessen et al., 1987). glands, adrenal glands, smooth muscle, prostate and eEF1G seems to have in human cells a dimeric urinary bladder. nature, forming homodimers, while in rabbit shows No protein presence is found in bone marrow, heart a trimeric nature and in yeast was observed that it muscle, kidney, liver and skeletal muscle (Fagerberg acts as a monomer (Achilonu et al.,2014; Koonin et et al., 2014). Furthermore was revealed the presence al., 1994). Furthermore, it has hydrophobic of the protein both in the human physiological properties that enable it to attach to membranes secretions (cerumen, saliva, milk, urine, seminal (Mansilla et al., 2002). plasma) and in pathological intercellular fluids eEF1G interacts mainly with EEF1B2 and EEF1D, (ascites) (https://www.proteomicsdb.org). even if other interactions are documented in protein databases and in literature, i.e. with the histidyl- tRNA synthetase (HARS), leucyl-tRNA synthetase

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Figure 3. Translation elongation mechanism. The active form of eukaryotic elongation factor 1 (eEF1A) in complex with GTP delivers an aminoacylated tRNA to the A site of the ribosome. Following the proper codon-anticodon recognition the GDP is hydrolyzed and the inactive eEF1A-GDP is released from the ribosome and then it is bound by eEF1B2GD complex forming the macromolecular protein aggregate eEF1H. eEF1H is formed previously by the binding of three subunits: eEF1B2, eEF1G and eEF1D. This complex promotes the exchange between GDP and GTP to regenerate active form of eEF1A (reworked from Dongsheng et al., 2013; Ejiri, 2002; Riis et al, 1990; https://reactome.org)

The expression pattern in cell lines tested for its the ribosome (Mansilla et al., 2002; Janssen et al., presence is similar and without significantly 1994). However, the complete significance of the differences except in one study that revealed a higher role of human eEF1G remains unknown and needs expression in human embryonic kidney HEK293 cell to be more studied. line and in human liver hepatocellular carcinoma Non-canonical roles: eEF1G has shown to interact HepG2 cell line (Cao et al, 2014). with cytoskeleton, RNA polymerase II, TNF Localisation receptor-associated protein 1 (TRAP1) and membrane-bound receptors. In addition, it was eEF1G is located mostly in the cytoplasm where observed that it has mRNA binding properties and it forming a gradient from the nucleus to the periphery is a positive regulator of NF-kB signaling pathway. of the cell, but some studies find it also in cellular 1) eEF1G and cytoskeleton: it was discovered that nucleus, nucleolus, mitochondria and in relation with eEF1G is a structural component of the cytoskeleton endoplasmic reticulum and plasma membrane (Cho (Coumans et al., 2014), in fact it can bind both et al., 2003; Minella et al., 1996; Sanders et al., keratin intermediate filaments (Kim et al., 2017) and 1996). It was reported also its localization in the tubulin (Janssen and Möller, 1988). This extracellular exosomes (Principe et al., 2013). suggests that it may have an influence on Function cytoskeletal architecture, cell morphology and eEF1G shows canonical functions and multiple non- motility, but these implications and the roles of these canonical roles (moonlighting roles) inside the cell. bindings are still need to clarify. Canonical function: eEF1G binds to eEF1B and 2) eEF1G and interaction with RNA polymerase eEF1D and is supposed that its primary role may be II: it physically interacts with RNA polymerase II to ensure the proper scaffolding and stability of its (pol II) core subunit 3 ( POLR2C), both in isolation binding partners in the eEF1BDG macromolecular and in the context of the holo- (Corbi et al., complex and then it could anchors the entire EF1H 2010). complex to the endoplasmic reticulum together with

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3) interaction between eEF1G and TNF The great number of mutations in the genomic receptor-associated protein 1 (TRAP1): TRAP1 sequence or in the amino acid sequence for EEF1G is the main mitochondrial member of the heat shock was discovered in cancer cells that are obviously protein (HSP) 90 family and it was revealed that genetically more unstable respect normal cells. The there is an interaction between this protein and some genomic alterations observed include the formation members of eEF1H complex, including eEF1G. of novel fusion genes as EBF3/EEF1G, The role of the interaction between eEF1G and EEF1G/ALK, EEF1G/ENG, EEF1G/GFAP, TRAP1 is related to the translational control EEF1G/MTA2, EEF1G/MYH9, EEF1G/NXF1, (Matassa et al., 2013) and maybe also in the EEF1G/OOEP [t(6;11)(q13;q12) EEF1G/OOEP], protection to oxidative stress (Pisani et al., 2016). EEF1G/PPP6R3 [t(11;11)(q12;q13) 4) eEF1G and membrane-bound receptors EEF1G/PPP6R3], EEF1G/TOX2, EEF1G/UBXN1, (dopamine D3 receptor): it was observed that there ETFB/EEF1G, HNRNPUL2/EEF1G, is an interaction between eEF1G and dopamine D3 IGHG1/EEF1G, IGHM/EEF1G and receptor ( DRD3) and that they have a co- NCEH1/EEF1G (Klijn et al., 2015; localization on the plasma membrane. http://atlasgeneticsoncology.org/Bands/11q12.html; This interaction involve also eEF1B subunit after its http://quiver.archerdx.com), however there are no protein kinase-mediated phosphorylation on its experimental data yet to understand the serine residues. repercussions on cellular behavior and so the eEF1G acts as a bridge for the relation between implications in cancer of these fusion genes are eEF1B and DRD3 and these three factors together unclear. forming a new macromolecular complex on the There is one chromosomic translocations with plasma membrane that obviously play some roles production of a novel fusion gene that is more even if its functional meaning is still understood investigated and it is t(2;11)(p23;q12.3) (Cho et al., 2003). EEF1G/ALK. 5) mRNA binding properties: the presence of Similarity Organism (1) Organism (2) Symbol eEF1G was detected on the genomic locus (%) corresponding to the promoter region of human Human H.sapiens eEF1G 100 vimentin gene VIM and this permits to suppose that Chimpanzee P.troglodytes eEF1G 100 eEF1G regulates vimentin gene by contacting both G.gorilla Gorilla eEF1G 99 pol II and the vimentin promoter region. gorilla In addition was shown that it can bind to 3'UTR of vimentin mRNA and so it can shuttling/nursing the Cat Felis catus eEF1G 99 vimentin mRNA from its gene locus to its Mouse M.musculus eEF1G 98 appropriate cellular compartment for translation. In Rat R.norvegicus eEF1G 98 fact, depletion of eEF1G causes the incorrect Zebrafish D.rerio eEF1G 75 compartmentalizing of the vimentin protein and Drosophila D.melanogaster Ef1gamma 58 seriously compromise cellular shape and Caenorhabditis C.elegans F17C11.9 52 mitochondria localization (Corbi et al., 2010). Furthermore, was shown that eEF1G can bind also Yeast S.cerevisiae TEF4 38 AATF (Che-1) and transcription and their promoter Table.3 eEF1G homology (reworked from regions (Pisani et al., 2016). https://cgap.nci.nih.gov; 6) regulation of NF-kB signaling pathway: eEF1G can binds to the CARD domain of Implicated in mitochondrial antiviral-signaling protein ( MAVS) Top note and thus significantly promotes the activities of High expression levels of eEF1G are observed in transcription factor NF-kB functioning as its positive many cancer types and, clinically, the regulator. overexpression of EEF1G was correlated with a poor The discover offers a new regulating mechanism of or better prognosis of cancers in relation to a specific the antiviral responses that promotes the downstream cancer type (Hassan et al., 2018). It is unclear if pro-inflammatory cytokines CXCL8 (interleukin-8 EFF1G overexpression concurs to the tumoral (IL8)) and interleukin-6 ( IL6) (Liu et al., 2014). process or it is a simply consequence and when it Homology occurs the mechanism of overexpression of EEF1G eEF1G is highly and abundant conserved in many is not known (Frazier et al., 1998). In addition, was species, with sometimes the lack of NT-eEF1G observed a few translocations with creation of fusion domain. The homology for eEF1G protein between genes with other proteins in some cancer cell types. species is reported in Table.3 Mutations

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t(2;11)(p23;q12.3) EEF1G/ ALK Breast Cancer Anaplastic lymphoma kinase (ALK)-positive High expression of EEF1G is observed in breast anaplastic large cell lymphoma (ALCL) is cancer cells and also in peripheral blood samples of characterized by many genomic aberrations and breast cancer patients respect to healthy subjects chromosomal rearrangements that make the cellular (Coumans et al., 2014 ; Aarøe et al., 2010), although caryotype much complicated. There are revealed other studies shown that it is down-regulated and many ALK aberrations and rearrangements with instead an increase of EEF1G transcript levels have several variant of partner fusion genes (van der positive correlation with distant metastasis free Krogt et al., 2017; van Krieken, 2017). survival (DMFS) and relapse free survival (RFS) and Prognosis so seems that over-expression of EEF1G is The prognosis is very poor, in fact patients correlated with a significantly good prognosis in expressing EEF1G/ALK fusion gene have shown an luminal A subtype (Hassan et al., 2018). EEF1G is unfavorable clinical course with fatal outcome. considered a negative marker for ERPR positive breast cancer (Tyanova et al., 2016). Hybrid/Mutated gene It was observed in some ALK+ ALCL pediatric Cervical carcinoma patients the presence of an in-frame fusion transcript EEF1G is observed to be highly expressed in between an intronic region among exons 8 and 9 of cervical intraepithelial neoplasia (Shadeo et al., EEF1G with the middle part of exon 20 of ALK. The 2008). resulting novel fusion chimeric gene 5'EEF1G / 3'ALK was revealed to be a coding-gene (van der Colorectal cancer Krogt et al., 2017). On the contrary, other authors EEF1G is over-expressed by twofold to tenfold in a found a fusion gene originated by the fusion of exon great percentage of colorectal adenomas and by 6 of EEF1G with the exon 20 of ALK (Palacios et twofold to 26-fold in the colorectal carcinoma al., 2017). compared to normal tissue and also its relative Abnormal protein protein eEF1G was found over-expressed, The chimeric protein eEF1G/ALK is active and has suggesting that early modification of its expression the complete GST-like N-terminal domain and part levels occurs and so it may be a useful marker for the of the CT domain of EEF1G fused to the complete detection of an early stage of tumor development intracellular protein tyrosine kinase (PTK) domain (Frazier et al., 1998; Ender et al., 1993; Chi et al., of ALK. Cytoplasm is the subcellular localization for 1992 ; Mathur et al., 1988). this chimera (van der Krogt et al., 2017; van Krieken, The overexpression of eEF1G in the colorectal 2017) but its biological activities, its oncogenic cancers seems to be not due to gene amplification, potential and its roles in proliferation and cancer genome rearrangements or an increase in the number aggressiveness are still poor understood although it of cycling cells (Frazier et al., 1998). is supposed that eEF1G/ALK fusion protein has a Interestingly is the positive correlation that was cell-transforming activity due to the activation of found in this cancer type between co-expression ALK kinase (Palacios et al., 2017). levels of TNF Receptor Associated Protein 1 (TRAP1) and eEF1G: the majority of the TRAP1- Acute myeloid leukemia (AML) positive tumors exhibit an upregulation of eEF1G Acute myeloid leukemia (AML) is the most common and, on the contrary, tumors with low expression of and severe form of acute leukemia diagnosed in TRAP1 also exhibit low levels of expression of adults. EEF1G was find over-expressed in AML-M1 eEF1G (Matassa et al., 2013). This evidence may and AML-M2 samples from adult patients have an interesting significance in the increase or (Handschuh et al, 2017). decrease of tumor aggressiveness and in the Brain and central nervous system development of new anti-cancer strategies. Moreover, the reduction of expression levels of (CNS) cancer eEF1G in high-risk patients can predict poor survival EEF1G mRNA levels are significantly upregulated (Hassan et al., 2018). in brain and CNS cancers. It is found over-expressed in glioblastoma and glioma, however higher levels Esophageal carcinoma of expression are found also in low-risk patients, so eEF1G is overexpressed in only a minimum part of this can predict favourable survival outcome (Hassan esophageal carcinoma tissues examined respect et al., 2018). On the contrary, another study observed normal counterpart and there is not any evidence a down-regulation of EEF1G in glioblastoma between its expression level and the growth rates of multiform (Vastrad and Vastrad, 2018). tumor. However, cancers over-expressing eEF1G show more aggressiveness and show a more metastatic

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behavior respect cancer cells that not overexpressing correlated with worse survival outcome in lung this gene. adenocarcinoma (Hassan et al., 2018). On the contrary, eEF1G is over-expressed in all In addition, from a preliminary study was observed esophageal cancer cell lines tested (Frazier et al., an EEF1G differential expression between stage I 1998; Mimori et al., 1996). from stage II lung squamous carcinoma: in the Gastric cancer second EEF1G has high expression levels and this may be one of the causes of the increase of grade of EEF1G was found significantly overexpressed in malignancy. low-grade gastric adenomas (Takenawa et al., 2004) However, other investigations are needed to confirm while on the contrary in another study was observed this preliminary evidence (Wang et al., 2018). that its expression level is down-regulated in gastric cancer cells and that higher levels of its expression Hybrid/Mutated gene could predict a poor overall survival (OS) and first In squamous cell carcinoma of the lung was progression (FP)(Hassan et al., 2018). Other authors discovered the fusion gene t(11;11)(q12;q13) found not only an overexpression of this gene in EEF1G/PPP6R3 (Hammerman et al., 2012). gastric carcinomas but also that tumors Lymphoma overexpressing EEF1G have more vascular EEF1G is significant overexpressed in Burkitt's permeation/angiogenesis than the others (Frazier et lymphoma and diffuse large B-Cell Lymphoma al., 1998; Mimori et al., 1995). This evidence is very (Hassan et al., 2018). significant and could be suppose that an Hybrid/Mutated gene overexpression of EEF1G may be compatible with In acute lymphoblastic leukemia/lymphoblastic more aggressiveness of the gastric cancer cells that lymphoma was discovered the fusion gene show a higher expression levels for this protein. t(6;11)(q13;q12) EEF1G/OOEP (Atak et al., 2013) Head and neck squamous cell and the presence of translocation t (2;11)(p23; q12.3) carcinoma EEF1G/ALK was observed in pediatric anaplastic mRNA levels of EEF1G are downregulated in tumor lymphoma kinase (ALK)-positive anaplastic large tissues than normal but in high-risk patients these cell lymphoma (ALCL). levels become significantly higher (Hassan et al., Melanoma 2018). EEF1G gene is relevantly co-expressed with other Kidney cancer genes in melanoma subtype 6 (Gan et al., 2018). EEF1G expression level is significantly upregulated Hybrid/Mutated gene in kidney clear cell carcinoma. 5'EEF1G /3'NXF1 fusion gene is observed in skin These high expression levels may predict better cutaneous melanoma survival in low-risk patients (Hassan et al., 2018). (https://www.empiregenomics.com). Liver cancer Multiple myeloma EEF1G is significant overexpressed in myeloma mRNA levels of EEF1G are significantly cells in the bone marrow of multiple myeloma upregulated in liver cancer cells respect normal ones patients (Sariman et al., 2019). and this may predict worse survival in high-risk patients (Hassan et al., 2018 ; Wang et al., 2009). Ovarian cancer In particular, mRNA expression levels of EEF1G High expression of EEF1G is found in ovarian remain at basal levels in a well to moderately cancer and this may predict a better overall survival differentiated (W/M-) primary human hepatocellular (OS) and progression-free survival (PFS)(Hassan et carcinoma (HCC), while they are further up- al., 2018). regulated in moderately to poorly differentiated Pancreatic cancer (M/P-) HCC according their histological grading (Shuda et al, 2000). From collected data seem to be no significantly In contrast, in vitro studies on cell cultures of HBV- difference between the expression levels of EEF1G or HCV-HCC have shown in most cases a down- in pancreatic cancer cells compared with normal regulation of EEF1G expression levels (Yoon et al., ones (Hassan et al., 2018), however in some studies 2006). were evidenced that EEF1G is over-expressed (Frazier et al., 1998 ; Chi et al., 1992 ; Lew et al., Lung cancer 1992). The presence of a higher level of its The expression levels of EEF1G are slightly higher expression may become a marker of poor in lung cancer cells respect normal ones and this lead survivability for high-risk patients (Hassan et al., to poor overall survival (OS) and first progression 2018). (FP) in lung cancer and thus are significantly

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EEF1G (Eukaryotic translation elongation factor 1 gamma) Cristiano L

Pleomorphic adenoma of the human Achilonu I, Siganunu TP, Dirr HW. Purification and characterisation of recombinant human eukaryotic parotid gland elongation factor 1 gamma. Protein Expr Purif. 2014 There is one study in which were found down- Jul;99:70-7 expressed levels for this gene (Mutlu et al., 2017). Atak ZK, Gianfelici V, Hulselmans G, De Keersmaecker K, Devasia AG, Geerdens E, Mentens N, Chiaretti S, Durinck Prostate cancer K, Uyttebroeck A, Vandenberghe P, Wlodarska I, Cloos J, Foà R, Speleman F, Cools J, Aerts S. Comprehensive A significant overexpression of EEF1G is seen in analysis of transcriptome variation uncovers known and prostate tumor tissues although this evidence seems novel driver events in T-cell acute lymphoblastic leukemia. not to affect the survival outcomes (Hassan et al., PLoS Genet. 2013;9(12):e1003997 2018). Remarkable is the discovery of presence of . Comprehensive genomic characterization of squamous eEF1G in exosomes contained in expressed prostatic cell lung cancers. Nature. 2012 Sep 27;489(7417):519-25 secretions (EPS) that could be utilized as diagnostic Cao Y, Portela M, Janikiewicz J, Doig J, Abbott CM. and/or prognostic markers for prostate cancer Characterisation of translation elongation factor eEF1B (Principe et al., 2013). subunit expression in mammalian cells and tissues and co- localisation with eEF1A2. PLoS One. 2014;9(12):e114117 To be noted Chi K, Jones DV, Frazier ML. Expression of an elongation factor 1 gamma-related sequence in adenocarcinomas of Roles of eEF1G in viral replication and the colon. Gastroenterology. 1992 Jul;103(1):98-102 pathogenesis Cho DI, Oak MH, Yang HJ, Choi HK, Janssen GM, Kim KM. eEF1G is an abundant cellular resource that seems to Direct and biochemical interaction between dopamine D3 play an important role in the genomic replication, receptor and elongation factor-1Bbetagamma. Life Sci. DNA synthesis, transcription and in the pathogenesis 2003 Oct 24;73(23):2991-3004 of a variety of viruses, such as HIV-1 and influenza Choudhary C, Kumar C, Gnad F, Nielsen ML, Rehman M, A virus, by interacting with viral polymerases, Walther TC, Olsen JV, Mann M. Lysine acetylation targets structural and nonstructural proteins, and viral protein complexes and co-regulates major cellular genome. functions. Science. 2009 Aug 14;325(5942):834-40 When eEF1G is downregulated using a small Corbi N, Batassa EM, Pisani C, Onori A, Di Certo MG, interfering RNA (siRNA) also the viral replication is Strimpakos G, Fanciulli M, Mattei E, Passananti C. The eEF1γ subunit contacts RNA polymerase II and binds negatively affected and becomes more unstable and vimentin promoter region. PLoS One. 2010 Dec less efficient. 31;5(12):e14481 However, the molecular interaction between the Coumans JV, Gau D, Poljak A, Wasinger V, Roy P, Moens virus components and eEF1G or other translation P. Green fluorescent protein expression triggers proteome factors remains to be determined (Sammaibashi et changes in breast cancer cells Exp Cell Res 2014 Jan al., 2018; Dongsheng et al., 2013 ; Warren et al., 1;320(1):33-45 2012). Ejiri S. Moonlighting functions of polypeptide elongation Reduction of cell viability after causing the factor 1: from actin bundling to zinc finger protein R1- down-regulation by RNAi associated nuclear localization Biosci Biotechnol Biochem 2002 Jan;66(1):1-21 The knockdown of eEF1G subunit by a specific siRNA shown a slightly reduced cell viability/cell Ender B, Lynch P, Kim YH, Inamdar NV, Cleary KR, Frazier metabolism. In addition, was observed that its ML. Overexpression of an elongation factor-1 gamma- hybridizing RNA in colorectal adenomas Mol Carcinog depletion can affect the expression of the eEF1B and 1993;7(1):18-20 eEF1D subunits (Cao et al., 2014). Fagerberg L, Hallström BM, Oksvold P, et al. Analysis of the Down-regulation of EEF1G in response to human tissue-specific expression by genome-wide enterovirus 71 (EV71) infection. integration of transcriptomics and antibody-based The transcriptomic and proteomic analyses of the proteomics Mol Cell Proteomics 2014 Feb;13(2):397-406 rhabdomyosarcoma cells infected by enterovirus 71 Frazier ML, Inamdar N, Alvula S, Wu E, Kim YH. Few point (EV71) has revealed a change in mutations in elongation factor-1gamma gene in profiles of several genes in response to infection. In gastrointestinal carcinoma Mol Carcinog 1998 May;22(1):9- particular, was observed the down-regulation of 15 EEF1G (Leong and Chow, 2006) although its Gan Y, Li N, Zou G, Xin Y, Guan J. Identification of cancer significance remain unclear. subtypes from single-cell RNA-seq data using a consensus clustering method BMC Med Genomics 2018 Dec References 31;11(Suppl 6):117 Han P, Chen C. Detergent-free biotin switch combined with Aarøe J, Lindahl T, Dumeaux V, Saebø S, Tobin D, Hagen liquid chromatography/tandem mass spectrometry in the N, Skaane P, Lönneborg A, Sharma P, Børresen-Dale AL. analysis of S-nitrosylated proteins Rapid Commun Mass Gene expression profiling of peripheral blood cells for early Spectrom 2008 Apr;22(8):1137-45 detection of breast cancer. Breast Cancer Res. 2010;12(1):R7 Handschuh L, Kamierczak M, Milewski MC, Góralski M, uczak M, Wojtaszewska M, Uszczyńska-Ratajczak

Atlas Genet Cytogenet Oncol Haematol. 2020; 24(2) 66

EEF1G (Eukaryotic translation elongation factor 1 gamma) Cristiano L

B, Lewandowski K, Komarnicki M, Figlerowicz M. Gene Maessen GD, Amons R, Zeelen JP, Möller W. Primary expression profiling of acute myeloid leukemia samples structure of elongation factor 1 gamma from Artemia FEBS from adult patients with AML-M1 and -M2 through boutique Lett 1987 Oct 19;223(1):181-6 microarrays, real-time PCR and droplet digital PCR Int J Oncol 2018 Mar;52(3):656-678 Mansilla F, Friis I, Jadidi M, Nielsen KM, Clark BF, Knudsen CR. Mapping the human translation elongation factor Hassan MK, Kumar D, Naik M, Dixit M. The expression eEF1H complex using the yeast two-hybrid system Biochem profile and prognostic significance of eukaryotic translation J 2002 Aug 1;365(Pt 3):669-76 elongation factors in different cancers PLoS One 2018 Jan 17;13(1):e0191377 Matassa DS, Amoroso MR, Agliarulo I, Maddalena F, Sisinni L, Paladino S, Romano S, Romano MF, Sagar V, Hirotsune S, Yoshida N, Chen A, Garrett L, Sugiyama F, Loreni F, Landriscina M, Esposito F. Translational control Takahashi S, Yagami K, Wynshaw-Boris A, Yoshiki A. An in the stress adaptive response of cancer cells: a novel role expressed pseudogene regulates the messenger-RNA for the heat shock protein TRAP1 Cell Death Dis 2013 Oct stability of its homologous coding gene Nature 2003 May 10;4:e851 1;423(6935):91-6 Mathur S, Cleary KR, Inamdar N, Kim YH, Steck P, Frazier Ishii H, Vecchione A, Murakumo Y, Baldassarre G, Numata ML. Overexpression of elongation factor-1gamma protein in S, Trapasso F, Alder H, Baffa R, Croce CM. FEZ1/LZTS1 colorectal carcinoma Cancer 1998 Mar 1;82(5):816-21 gene at 8p22 suppresses cancer cell growth and regulates Mimori K, Mori M, Tanaka S, Akiyoshi T, Sugimachi K. The mitosis Proc Natl Acad Sci U S A 2001 Aug overexpression of elongation factor 1 gamma mRNA in 28;98(18):10374-9 gastric carcinoma Cancer 1995 Mar 15;75(6 Suppl):1446- 9 Janssen GM, van Damme HT, Kriek J, Amons R, Möller W. The subunit structure of elongation factor 1 from Artemia Minella O, Mulner-Lorillon O, De Smedt V, Hourdez S, Why two alpha-chains in this complex? J Biol Chem 1994 Cormier P, Bellé R. Major intracellular localization of Dec 16;269(50):31410-7 elongation factor-1 Cell Mol Biol (Noisy-le-grand) 1996 Sep;42(6):805-10 Jiang S, Wolfe CL, Warrington JA, Norcum MT. Three- dimensional reconstruction of the valyl-tRNA Mutlu A, Ozturk M, Akpinar G, Kasap M, Kanli A. Proteomics synthetase/elongation factor-1H complex and localization of analysis of pleomorphic adenoma of the human parotid the delta subunit FEBS Lett 2005 Nov 7;579(27):6049-54 gland Eur Arch Otorhinolaryngol 2017 Aug;274(8):3183- 3195 Kim S, Kellner J, Lee CH, Coulombe PA. Interaction between the keratin cytoskeleton and eEF1Bgamma affects Olsen JV, Vermeulen M, Santamaria A, Kumar C, Miller ML, protein synthesis in epithelial cells Nat Struct Mol Biol 2007 Jensen LJ, Gnad F, Cox J, Jensen TS, Nigg EA, Brunak S, Oct;14(10):982-3 Mann M. Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during Klijn C, Durinck S, Stawiski EW, Haverty PM, Jiang Z, Liu mitosis Sci Signal 2010 Jan 12;3(104):ra3 H, Degenhardt J, Mayba O, Gnad F, Liu J, Pau G, Reeder J, Cao Y, Mukhyala K, Selvaraj SK, Yu M, Zynda GJ, Brauer Ota T, Suzuki Y, Nishikawa T, et al. Complete sequencing MJ, Wu TD, Gentleman RC, Manning G, Yauch RL, and characterization of 21,243 full-length human cDNAs Bourgon R, Stokoe D, Modrusan Z, Neve RM, de Sauvage Nat Genet 2004 Jan;36(1):40-5 FJ, Settleman J, Seshagiri S, Zhang Z. A comprehensive transcriptional portrait of human cancer cell lines Nat Palacios G, Shaw TI, Li Y, Singh RK, Valentine M, Sandlund Biotechnol 2015 Mar;33(3):306-12 JT, Lim MS, Mullighan CG, Leventaki V. Novel ALK fusion in anaplastic large cell lymphoma involving EEF1G, a Koonin EV, Mushegian AR, Tatusov RL, Altschul SF, Bryant subunit of the eukaryotic elongation factor-1 complex SH, Bork P, Valencia A. Eukaryotic translation elongation Leukemia 2017 Mar;31(3):743-747 factor 1 gamma contains a glutathione transferase domain- -study of a diverse, ancient protein superfamily using motif Pisani C, Onori A, Gabanella F, Delle Monache F, Borreca search and structural modeling Protein Sci 1994 A, Ammassari-Teule M, Fanciulli M, Di Certo MG, Nov;3(11):2045-54 Passananti C, Corbi N. eEF1Bγ binds the Che-1 and TP53 gene promoters and their transcripts J Exp Clin Cancer Res Le Sourd F, Boulben S, Le Bouffant R, Cormier P, Morales 2016 Sep 17;35(1):146 J, Belle R, Mulner-Lorillon O. eEF1B: At the dawn of the 21st century Biochim Biophys Acta 2006 Jan-Feb;1759(1- Principe S, Jones EE, Kim Y, Sinha A, Nyalwidhe JO, 2):13-31 Brooks J, Semmes OJ, Troyer DA, Lance RS, Kislinger T, Drake RR. In-depth proteomic analyses of exosomes Leong WF, Chow VT. Transcriptomic and proteomic isolated from expressed prostatic secretions in urine analyses of rhabdomyosarcoma cells reveal differential Proteomics 2013 May;13(10-11):1667-1671 cellular gene expression in response to enterovirus 71 infection Cell Microbiol 2006 Apr;8(4):565-80 Riis B, Rattan SI, Clark BF, Merrick WC. Eukaryotic protein elongation factors Trends Biochem Sci 1990 Lew Y, Jones DV, Mars WM, Evans D, Byrd D, Frazier ML. Nov;15(11):420-4 Expression of elongation factor-1 gamma-related sequence in human pancreatic cancer Pancreas 1992;7(2):144-52 Sammaibashi S, Yamayoshi S, Kawaoka Y. Strain-Specific Contribution of Eukaryotic Elongation Factor 1 Gamma to Li D, Wei T, Abbott CM, Harrich D. The unexpected roles of the Translation of Influenza A Virus Proteins Front Microbiol eukaryotic translation elongation factors in RNA virus 2018 Jun 29;9:1446 replication and pathogenesis Microbiol Mol Biol Rev 2013 Jun;77(2):253-66 Sanders J, Brandsma M, Janssen GM, Dijk J, Möller W. Immunofluorescence studies of human fibroblasts Liu D, Sheng C, Gao S, Jiang W, Li J, Yao C, Chen H, Wu demonstrate the presence of the complex of elongation J, Chen S, Huang W. eEF1Bγ is a positive regulator of NF- factor-1 beta gamma delta in the endoplasmic reticulum J B signaling pathway Biochem Biophys Res Commun 2014 Cell Sci 1996 May;109 ( Pt 5):1113-7 Apr 4;446(2):523-8

Atlas Genet Cytogenet Oncol Haematol. 2020; 24(2) 67

EEF1G (Eukaryotic translation elongation factor 1 gamma) Cristiano L

Sariman M, Abaci N, Sirma Ekmekçi S, akiris A, Perçin Gastroenterol Hepatol 2009 Apr;24(4):605-17 Paçal F, Üstek D, Ayer M, Yenerel MN, Bek S, efle K, Palandüz , Öztürk. Investigation of Gene Expressions of Wang R, Cai Y, Zhang B, Wu Z. A 16-gene expression Myeloma Cells in the Bone Marrow of Multiple Myeloma signature to distinguish stage I from stage II lung squamous Patients by Transcriptome Analysis Balkan Med J 2019 Jan carcinoma Int J Mol Med 2018 Mar;41(3):1377-1384 1;36(1):23-31 Warren K, Wei T, Li D, Qin F, Warrilow D, Lin MH, Shadeo A, Chari R, Lonergan KM, Pusic A, Miller D, Ehlen Sivakumaran H, Apolloni A, Abbott CM, Jones A, Anderson T, Van Niekerk D, Matisic J, Richards-Kortum R, Follen M, JL, Harrich D. Eukaryotic elongation factor 1 complex Guillaud M, Lam WL, MacAulay C. Up regulation in gene subunits are critical HIV-1 reverse transcription cofactors expression of chromatin remodelling factors in cervical Proc Natl Acad Sci U S A 2012 Jun 12;109(24):9587-92 intraepithelial neoplasia BMC Genomics 2008 Feb 4;9:64 Yoon SY, Kim JM, Oh JH, Jeon YJ, Lee DS, Kim JH, Choi Sheu GT, Traugh JA. A structural model for elongation JY, Ahn BM, Kim S, Yoo HS, Kim YS, Kim NS. Gene factor 1 (EF-1) and phosphorylation by protein kinase CKII expression profiling of human HBV- and/or HCV-associated Mol Cell Biochem 1999 Jan;191(1-2):181-6 hepatocellular carcinoma cells using expressed sequence Shuda M, Kondoh N, Tanaka K, Ryo A, Wakatsuki T, Hada tags Int J Oncol 2006 Aug;29(2):315-27 A, Goseki N, Igari T, Hatsuse K, Aihara T, Horiuchi S, van Damme H, Amons R, Janssen G, Möller W. Mapping Shichita M, Yamamoto N, Yamamoto M. Enhanced the functional domains of the eukaryotic elongation factor 1 expression of translation factor mRNAs in hepatocellular beta gamma Eur J Biochem 1991 Apr 23;197(2):505-11 carcinoma Anticancer Res 2000 Jul-Aug;20(4):2489-94 van Krieken JH. New developments in the pathology of Takenawa H, Kurosaki M, Enomoto N, Miyasaka Y, malignant lymphoma: a review of the literature published Kanazawa N, Sakamoto N, Ikeda T, Izumi N, Sato C, from May to August 2017 J Hematop 2017 Sep Watanabe M. Differential gene-expression profiles 30;10(2):65-73 associated with gastric adenoma Br J Cancer 2004 Jan 12;90(1):216-23 van der Krogt JA, Bempt MV, Ferreiro JF, Mentens N, Jacobs K, Pluys U, Doms K, Geerdens E, Uyttebroeck A, Tyanova S, Albrechtsen R, Kronqvist P, Cox J, Mann M, Pierre P, Michaux L, Devos T, Vandenberghe P, Tousseyn Geiger T. Proteomic maps of breast cancer subtypes Nat T, Cools J, Wlodarska I. Anaplastic lymphoma kinase- Commun 2016 Jan 4;7:10259 positive anaplastic large cell lymphoma with the variant Vastrad C, Vastrad B. Bioinformatics analysis of gene RNF213-, ATIC- and TPM3-ALK fusions is characterized by expression profiles to diagnose crucial and novel genes in copy number gain of the rearranged ALK gene glioblastoma multiform Pathol Res Pract 2018 Haematologica 2017 Sep;102(9):1605-1616 Sep;214(9):1395-1461 This article should be referenced as such: Wang F, Kuang Y, Salem N, Anderson PW, Lee Z. Cross- Cristiano L. EEF1G (Eukaryotic translation elongation species hybridization of woodchuck hepatitis viral infection- factor 1 gamma). Atlas Genet Cytogenet Oncol induced woodchuck hepatocellular carcinoma using human, Haematol. 2020; 24(2):58-68. rat and mouse oligonucleotide microarrays J

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