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Nahum Sonenberg - One of the best experts on this subject based on the ideXlab platform.
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a novel function of the ma 3 domains in transformation and translation suppressor pdcd4 is essential for its binding to eukaryotic translation Initiation Factor 4A
Molecular and Cellular Biology, 2004Co-Authors: Hsinsheng Yang, Nahum Sonenberg, Myunghaing Cho, Halina Zakowicz, Glenn Hegamyer, Nancy H ColburnAbstract:Αn α-helical MA-3 domain appears in several translation Initiation Factors, including human eukaryotic translation Initiation Factor 4G (eIF4G) and DAP-5/NAT1/p97, as well as in the tumor suppressor Pdcd4. The function of the MA-3 domain is, however, unknown. C-terminal eIF4G (eIG4Gc) contains an MA-3 domain that is located within the eIF4A-binding region, suggesting a role for eIF4A binding. Interestingly, C-terminal DAP-5/NAT1/p97 contains an MA-3 domain, but it does not bind to eIF4A. Mutation of amino acid residues conserved between Pdcd4 and eIF4Gc but not in DAP-5/NAT1/p97 to the amino acid residues found in the DAP-5/NAT1/p97 indicates that some of these amino acid residues within the MA-3 domain are critical for eIF4A-binding activity. Six Pdcd4 mutants (Pdcd4E249K, Pdcd4D253A, Pdcd4D414K, Pdcd4D418A, Pdcd4E249K,D414K, and Pdcd4D253A,D418A) lost >90% eIF4A-binding activity. Mutation of the corresponding amino acid residues in the eIF4Gc also produced similar results, as seen for Pdcd4. These results demonstrate that the MA-3 domain is important for eIF4A binding and explain the ability of Pdcd4 or eIF4Gc but not DAP-5/NAT1/p97 to bind to eIF4A. Competition experiments indicate that Pdcd4 prevents ca. 60 to 70% of eIF4A binding to eIF4Gc at a Pdcd4/eIF4A ratio of 1:1, but mutants Pdcd4D253A and Pdcd4D253A,D418A do not. Translation of stem-loop structured mRNA is susceptible to inhibition by wild-type Pdcd4 but not by Pdcd4D253A, Pdcd4D418A, or Pdcd4D235A,D418A. Together, these results indicate that not only binding to eIF4A but also prevention of eIF4A binding to the MA-3 domain of eIF4Gc contributes to the mechanism by which Pdcd4 inhibits translation.
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a nuclear translation like Factor eif4Aiii is recruited to the mrna during splicing and functions in nonsense mediated decay
Proceedings of the National Academy of Sciences of the United States of America, 2004Co-Authors: Maria Ferraiuolo, Chungsheng Lee, Lian Wee Ler, Jeanne L Hsu, Mauro Costamattioli, Mingjuan Luo, Robin Reed, Nahum SonenbergAbstract:In eukaryotes, a surveillance mechanism known as nonsense-mediated decay (NMD) degrades the mRNA when a premature-termination codon (PTC) is present. NMD requires translation to read the frame of the mRNA and detect the PTC. During pre-mRNA splicing, the exon–exon junction complex (EJC) is recruited to a region 20–24 nt upstream of the exon junction on the mature mRNA. The presence of a PTC upstream from the EJC elicits NMD. Eukaryotic Initiation Factor 4A (eIF4A) III is a nuclear protein that interacts physically or functionally with translation Initiation Factors eIF4G and eIF4B, respectively, and shares strikingly high identity with the Initiation Factors eIF4AI/II. Here we show that siRNA against eIF4AIII, but not against eIF4AI/II, inhibits NMD. Moreover, eIF4AIII, but not eIF4AI, is specifically recruited to the EJC during splicing. The observations that eIF4AIII is loaded onto the mRNA during splicing in the nucleus, has properties related to a translation Initiation Factor, and functions in NMD raises the possibility that eIF4AIII substitutes for eIF4AI/II during NMD.
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eIF4AIII binds spliced mRNA in the exon junction complex and is essential for nonsense-mediated decay
Nature Structural and Molecular Biology, 2004Co-Authors: Toshiharu Shibuya, Thomas Tange, Nahum Sonenberg, Melissa J MooreAbstract:The exon junction complex (EJC), a set of proteins deposited on mRNAs as a consequence of pre-mRNA splicing, is a key effector of downstream mRNA metabolism. We have identified eIF4AIII, a member of the eukaryotic translation Initiation Factor 4A family of RNA helicases (also known as DExH/D box proteins), as a novel EJC core component. Crosslinking and antibody inhibition studies suggest that eIF4AIII constitutes at least part of the platform anchoring other EJC components to spliced mRNAs. A nucleocytoplasmic shuttling protein, eIF4AIII associates in vitro and in vivo with two other EJC core Factors, Y14 and Magoh. In mammalian cells, eIF4AIII is essential for nonsense-mediated mRNA decay (NMD). Finally, a model is proposed by which eIF4AIII represents a new functional class of DExH/D box proteins that act as RNA clamps or 'place holders' for the sequence-independent attachment of additional Factors to RNAs.
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rna aptamers to Initiation Factor 4A helicase hinder cap dependent translation by blocking atp hydrolysis
RNA, 2003Co-Authors: Akihiro Oguro, Nahum Sonenberg, Takashi Ohtsu, Yuri V Svitkin, Yoshikazu NakamuraAbstract:The mammalian translation Initiation Factor 4A (eIF4A) is a prototype member of the DEAD-box RNA helicase family that couples ATPase activity to RNA binding and unwinding. In the crystal form, eIF4A has a distended ‘‘dumbbell’’ structure consisting of two domains, which probably undergo a conformational change, on binding ATP, to form a compact, functional structure via the juxtaposition of the two domains. Moreover, additional conformational changes between two domains may be involved in the ATPase and helicase activity of eIF4A. The molecular basis of these conformational changes, however, is not understood. Here, we generated RNA aptamers with high affinity for eIF4A by in vitro RNA selection-amplification. On binding, the RNAs inhibit ATP hydrolysis. One class of RNAs contains members that exhibit dissociation constant of 27 nM for eIF4A and severely inhibit cap-dependent in vitro translation. The binding affinity was increased on Arg substitution in the conserved motif Ia of eIF4A, which probably improves a predicted arginine network to bind RNA substrates. Selected RNAs, however, failed to bind either domain of eIF4A that had been split at the linker site. These findings suggest that the selected RNAs interact cooperatively with both domains of eIF4A, either in the dumbbell or the compact form, and entrap it into a dead-end conformation, probably by blocking the conformational change of eIF4A. The selected RNAs, therefore, represent a new class of specific inhibitors that are suitable for the analysis of eukaryotic Initiation, and which pose a potential therapeutic against malignancies that are caused by aberrant translational control.
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the transformation suppressor pdcd4 is a novel eukaryotic translation Initiation Factor 4A binding protein that inhibits translation
Molecular and Cellular Biology, 2003Co-Authors: Hsinsheng Yang, William C. Merrick, Nahum Sonenberg, Aaron P Jansen, Anton A Komar, Xiaojing Zheng, Sylvain V Costes, Stephen J Lockett, Nancy H ColburnAbstract:Pdcd4 is a novel transformation suppressor that inhibits tumor promoter-induced neoplastic transformation and the activation of AP-1-dependent transcription required for transformation. A yeast two-hybrid analysis revealed that Pdcd4 associates with the eukaryotic translation Initiation Factors eIF4AI and eIF4AII. Immunofluorescent confocal microscopy showed that Pdcd4 colocalizes with eIF4A in the cytoplasm. eIF4A is an ATP-dependent RNA helicase needed to unwind 5' mRNA secondary structure. Recombinant Pdcd4 specifically inhibited the helicase activity of eIF4A and eIF4F. In vivo translation assays showed that Pdcd4 inhibited cap-dependent but not internal ribosome entry site (IRES)-dependent translation. In contrast, Pdcd4(D418A), a mutant inactivated for binding to eIF4A, failed to inhibit cap-dependent or IRES-dependent translation or AP-1 transactivation. Recombinant Pdcd4 prevented eIF4A from binding to the C-terminal region of eIF4G (amino acids 1040 to 1560) but not to the middle region of eIF4G(amino acids 635 to 1039). In addition, both Pdcd4 and Pdcd4(D418A) bound to the middle region of eIF4G. The mechanism by which Pdcd4 inhibits translation thus appears to involve inhibition of eIF4A helicase, interference with eIF4A association-dissociation from eIF4G, and inhibition of eIF4A binding to the C-terminal domain of eIF4G. Pdcd4 binding to eIF4A is linked to its transformation-suppressing activity, as Pdcd4-eIF4A binding and consequent inhibition of translation are required for Pdcd4 transrepression of AP-1.
Narendra Tuteja - One of the best experts on this subject based on the ideXlab platform.
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pea dna helicase 45 promotes salinity stress tolerance in ir64 rice with improved yield
Plant Signaling & Behavior, 2012Co-Authors: Ranjan Kumar Sahoo, Sarvajeet Singh Gill, Narendra TutejaAbstract:The helicases provide duplex unwinding function in an ATP-dependent manner and thereby play important role in almost all the nucleic acids transaction. Since stress reduces the protein synthesis by affecting the cellular gene expression machinery, so it is evident that molecules involved in nucleic acid processing including translation Factors/helicases are likely to be affected. Earlier pea DNA helicase 45 (PDH45), a homolog of translation Initiation Factor 4A (eIF4A) was reported to play important role in salinity stress tolerance in tobacco and Bangladeshi rice variety Binnatoa. We report here the overexpression of PDH45 gene in the indica rice variety IR64, via Agrobacterium-mediated transformation. Molecular analysis of the transgenics revealed stable integration of the transgene in the T1 generation. Enhanced tolerance to salinity was observed in the plants transformed with PDH45 gene. Better physiological and yield performances including endogenous nutrient contents (N, P, K, Na) of the transgenics under salt treatment were observed as compared with wild type (WT), vector control and antisense transgenics. All these results indicated that the overexpression of PDH45 in the IR64 rice transgenics enable them to perform better with enhanced salinity stress tolerance and improved physiological traits. Based on the homology of PDH45 protein with eIF4A protein we suggest that it may act at the translational level to enhance or stabilize protein synthesis under stress conditions.
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pea dna helicase 45 promotes salinity stress tolerance in ir64 rice with improved yield
Plant Signaling & Behavior, 2012Co-Authors: Ranjan Kumar Sahoo, Sarvajeet Singh Gill, Narendra TutejaAbstract:The helicases provide duplex unwinding function in an ATP-dependent manner and thereby play important role in almost all the nucleic acids transaction. Since stress reduces the protein synthesis by affecting the cellular gene expression machinery, so it is evident that molecules involved in nucleic acid processing including translation Factors/helicases are likely to be affected. Earlier pea DNA helicase 45 (PDH45), a homolog of translation Initiation Factor 4A (eIF4A) was reported to play important role in salinity stress tolerance in tobacco and Bangladeshi rice variety Binnatoa. We report here the overexpression of PDH45 gene in the indica rice variety IR64, via Agrobacterium-mediated transformation. Molecular analysis of the transgenics revealed stable integration of the transgene in the T1 generation. Enhanced tolerance to salinity was observed in the plants transformed with PDH45 gene. Better physiological and yield performances including endogenous nutrient contents (N, P, K, Na) of the transgenics...
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Translation Initiation Factor 4A: a prototype member of dead-box protein family.
Physiology and Molecular Biology of Plants, 2008Co-Authors: Narendra Tuteja, Ajay A. Vashisht, Renu TutejaAbstract:DEAD-box proteins are characterized by nine conserved helicase motifs. Several different DEAD-box proteins are found in eukaryotes, whereas prokaryotes have small number of these proteins. They play important roles in almost all kinds of RNA metabolism including roles in remodeling ribonuclear protein complexes. These proteins are usually very specific and cannot mutually be replaced. Many of these DEAD-box proteins (but not all) have been shown to have RNA-dependent ATPase and ATP-dependent RNA helicase activities. Many of them have also been shown to contain DNA unwinding activities. Translation Initiation Factor 4A is the prototype of the DEAD-box family of proteins. Actually, the DEAD-box protein family was discovered on the basis of conserved sequence motifs of eIF4A. Motif II (or Walker B motif) has the amino acids D-E-A-D (Asp-Glu-Ala-Asp), which gave the name to the family. In general, the eIF4A is considered as a helicase that locally melts the secondary structures and makes the RNA accessible to nucleases. It is part of the cap-binding complex eIF4F but is also found to be present in a free form. The biochemical activities of eIF4A are reported to be upregulated by eIF4B, eIF4H and eIF4G. It has been proposed that eIF4A helps to unwind secondary structures in the 5′-untranslated region, which are inhibitory for protein synthesis. In plants, it has been shown to play a unique role in abiotic stress tolerance, which suggests a new pathway to engineer to increase the crop production under the stress conditions.
Jerry Pelletier - One of the best experts on this subject based on the ideXlab platform.
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intercepted retro nazarov reaction syntheses of amidino rocaglate derivatives and their biological evaluation as eif4A inhibitors
Journal of the American Chemical Society, 2019Co-Authors: Wenhan Zhang, Jerry Pelletier, Jennifer Chu, Andrew Cyr, Han Yueh, Lauren E Brown, Tony T Wang, John A PorcoAbstract:Rocaglates are a family of natural products isolated from the genus Aglaia which possess a highly substituted cyclopenta[b]benzofuran skeleton and inhibit cap-dependent protein synthesis. Rocaglates are attractive compounds due to their potential for inhibiting tumor cell maintenance in vivo by specifically targeting eukaryotic Initiation Factor 4A (eIF4A) and interfering with recruitment of ribosomes to mRNA. In this paper, we describe an intercepted retro-Nazarov reaction utilizing intramolecular tosyl migration to generate a reactive oxyallyl cation on the rocaglate skeleton. Trapping of the oxyallyl cation with a diverse range of nucleophiles has been used to generate over 50 novel amidino-rocaglate (ADR) and amino-rocaglate derivatives. Subsequently, these derivatives were evaluated for their ability to inhibit cap-dependent protein synthesis where they were found to outperform previous lead compounds including the rocaglate hydroxamate CR-1-31-B.
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hippuristanol a potent steroid inhibitor of eukaryotic Initiation Factor 4A
Translation (Austin Tex.), 2016Co-Authors: Regina Cencic, Jerry PelletierAbstract:Protein synthesis and its regulatory signaling pathways play essential roles in the Initiation and maintenance of the cancer phenotype. Insight obtained over the last 3 decades on the mechanisms regulating translation in normal and transformed cells have revealed that perturbed control in cancer cells may offer an Achilles' heel for the development of novel anti-neoplastic agents. Several small molecule inhibitors have been identified and characterized that target translation Initiation - more specifically, the rate-limiting step where ribosomes are recruited to mRNA templates. Among these, hippuristanol, a polyhydroxysteroid from the gorgonian Isis hippuris has been found to inhibit translation Initiation by blocking the activity of eukaryotic Initiation Factor (eIF) 4A, an essential RNA helicase involved in this process. Herein, we highlight the biological properties of this compound, its potential development as an anti-cancer agent, and its use to validate eIF4A as an anti-neoplastic target.
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Inhibitors of translation targeting eukaryotic translation Initiation Factor 4A.
Methods in enzymology, 2012Co-Authors: Regina Cencic, Gabriela Galicia-vázquez, Jerry PelletierAbstract:The RNA helicases eIF4AI and eIF4AII play key roles in recruiting ribosomes to mRNA templates during eukaryotic translation Initiation. Small molecule inhibitors of eIF4AI and eIF4AII have been useful for chemically dissecting their role in translation in vitro and in vivo. Here, we describe a screen performed on a small focused library of kinase inhibitors to identify a novel helicase inhibitor. We describe assays that have been critical for characterizing novel RNA helicase inhibitors.
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rna mediated sequestration of the rna helicase eif4A by pateamine a inhibits translation Initiation
Chemistry & Biology, 2006Co-Authors: Marieeve Bordeleau, Regina Cencic, Lisa M Lindqvist, Monika Oberer, Peter T Northcote, Gerhard Wagner, Jerry PelletierAbstract:Summary Eukaryotic Initiation Factor 4A (eIF4A) is a member of the DEAD-box family of putative RNA helicases whose members are involved in many aspects of RNA metabolism. eIF4A is thought to facilitate binding of 43S preInitiation complexes to mRNAs by unwinding secondary structures present in the 5′ untranslated region. Pateamine A, a small-molecule inhibitor of translation Initiation, acts in an unusual manner by stimulating eIF4A activity. Herein, we report the elucidation of pateamine's mode of action. We demonstrate that Pateamine A is a chemical inducer of dimerization that forces an engagement between eIF4A and RNA and prevents eIF4A from participating in the ribosome-recruitment step of translation Initiation.
Yan Lin - One of the best experts on this subject based on the ideXlab platform.
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eukaryotic Initiation Factor 4A 3 a review of its physiological role and involvement in oncogenesis
Frontiers in Oncology, 2021Co-Authors: Xiaomin She, Xing Gao, Rong Liang, Ziyu Liu, Yan LinAbstract:EIF4A3, a member of the DEAD-box protein family, is a nuclear matrix protein and a core component of the exon junction complex (EJC). Under physiological conditions, EIF4A3 participates in post-transcriptional gene regulation by promoting EJC control of precursor mRNA splicing, thus influencing nonsense-mediated mRNA decay. In addition, EIF4A3 maintains the expression of significant selenoproteins, including phospholipid hydroperoxide glutathione peroxidase and thioredoxin reductase 1. Several recent studies have shown that EIF4A3 promotes tumor growth in multiple human cancers such as glioblastoma, hepatocellular carcinoma, pancreatic cancer, and ovarian cancer. Molecular biology studies also showed that EIF4A3 is recruited by long non-coding RNAs to regulate the expression of certain proteins in tumors. However, its tumor-related functions and underlying mechanisms are not well understood. Here, we review the physiological role of EIF4A3 and the potential association between EIF4A3 overexpression and tumorigenesis. We also evaluate the protein's potential utility as a diagnosis biomarker, therapeutic target, and prognosis indicator, hoping to provide new ideas for future research.
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comprehensive analysis of biological networks and the eukaryotic Initiation Factor 4A 3 gene as pivotal in hepatocellular carcinoma
Journal of Cellular Biochemistry, 2020Co-Authors: Yan Lin, Rong Liang, Yingwei Mao, Rongyun Mai, Xing Gao, Ziyu Liu, Taylor Wainwright, Min Luo, Donghua ZouAbstract:Eukaryotic Initiation Factor 4A-3 (EIF4A3) is a core component of the exon junction complex (EJC). Abnormalities in EIF4A3 are associated with carcinogenesis. The present study aimed to determine the biological role of EIF4A3 in hepatocellular carcinoma (HCC). Our study is based on the analysis of HCC sequencing data from public databases. We first used the Gene Expression Profiling Interactive Analysis tool and ONCOMINE to analyze the EIF4A3 expression, and the results were validated in human clinical tissues by a quantitative real-time polymerase chain reaction, Western blot, and immunohistochemical. Then, we used cBioPortal to identify EIF4A3 alterations and function networks. Finally, we created a network of genes that were positively correlated with EIF4A3 using LinkedOmics, and analyzed this network using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. For the genes identified, we also analyzed the relevant kinase and transcription Factor target networks as well as the protein-protein interaction networks. Our results show that EIF4A3 was overexpressed in HCC tissues in comparison with normal tissues, and high EIF4A3 expression was significantly associated with poor prognosis. Analysis of the functional networks of genes that were co-occurring with EIF4A3 amplification revealed connections with several chemokine signaling pathways. Furthermore, genes that positively correlated with EIF4A3 were mainly related to cell cycle and spliceosome pathways, several cell cycle regulatory kinases, and tumor-associated transcription Factors. Finally, crosslinking-immunoprecipitation and high-throughput sequencing (CLIP-seq) data showed that EIF4A3 protein binds to multiple exon regions of the cell cycle regulatory genes cyclin-dependent kinases 1 and 2 and transcription Factor E2F1. Our study unveils potential biological networks in HCC and the pivotal role of EIF4A3 as a bridging protein, highlighting the need for an in-depth study of EIF4A3 in carcinogenesis.
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systematic analysis of gene expression alteration and co expression network of eukaryotic Initiation Factor 4A 3 in cancer
Journal of Cancer, 2018Co-Authors: Yan Lin, Jinyan Zhang, Junying Cai, Rong Liang, Guoying Chen, Gang Qin, Xueqiong Han, Chunling Yuan, Zhihui Liu, Donghua ZouAbstract:Purpose: Eukaryotic Initiation Factor 4A-3 (EIF4A3) is an RNA-binding protein (RBP) that is a core component of the exon junction complex (EJC). It has been identified as an important player in post-transcriptional regulation processes. Recently, investigations have focused on EIF4A3 dysfunction in carcinogenesis. The present study aims to determine whether EIF4A3 can serve as a prognostic marker and potential regulatory mechanism in human cancers. Materials and methods: EIF4A3 expression in various cancers was assessed using Oncomine. The Correlation between EIF4A3 expression and patient survival was evaluated using PrognoScan. EIF4A3 mutations in various cancers were investigated using cBioPortal. EIF4A3 co-expression networks in various cancers were established using Coexpedia. Finally, we analyzed potential functional roles of EIF4A3 using Gene Ontology and pathway enrichment analyses by FunRich V3. Results: EIF4A3 was overexpressed in common malignancies at the transcription levels. High incidences of the breast, lung, and urinary cancers were closely related to the prognostic index for survival. The most prevalent mutation in EIF4A3 was E59K/Q. The tumor necrosis Factor-α (TNF-α)/nuclear Factor-κB (NF-κB) signaling pathway was affected by these mutations. Co-expression networks showed that EIF4A3 regulates apoptosis and cell cycle via several cancer-related signal pathways, and promotes tumor cell migration, invasion and drug resistance. Conclusion: Our results suggest the potential role for EIF4A3 to serve as a diagnostic marker or therapeutic target for certain types of cancers.
Nancy H Colburn - One of the best experts on this subject based on the ideXlab platform.
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a novel function of the ma 3 domains in transformation and translation suppressor pdcd4 is essential for its binding to eukaryotic translation Initiation Factor 4A
Molecular and Cellular Biology, 2004Co-Authors: Hsinsheng Yang, Nahum Sonenberg, Myunghaing Cho, Halina Zakowicz, Glenn Hegamyer, Nancy H ColburnAbstract:Αn α-helical MA-3 domain appears in several translation Initiation Factors, including human eukaryotic translation Initiation Factor 4G (eIF4G) and DAP-5/NAT1/p97, as well as in the tumor suppressor Pdcd4. The function of the MA-3 domain is, however, unknown. C-terminal eIF4G (eIG4Gc) contains an MA-3 domain that is located within the eIF4A-binding region, suggesting a role for eIF4A binding. Interestingly, C-terminal DAP-5/NAT1/p97 contains an MA-3 domain, but it does not bind to eIF4A. Mutation of amino acid residues conserved between Pdcd4 and eIF4Gc but not in DAP-5/NAT1/p97 to the amino acid residues found in the DAP-5/NAT1/p97 indicates that some of these amino acid residues within the MA-3 domain are critical for eIF4A-binding activity. Six Pdcd4 mutants (Pdcd4E249K, Pdcd4D253A, Pdcd4D414K, Pdcd4D418A, Pdcd4E249K,D414K, and Pdcd4D253A,D418A) lost >90% eIF4A-binding activity. Mutation of the corresponding amino acid residues in the eIF4Gc also produced similar results, as seen for Pdcd4. These results demonstrate that the MA-3 domain is important for eIF4A binding and explain the ability of Pdcd4 or eIF4Gc but not DAP-5/NAT1/p97 to bind to eIF4A. Competition experiments indicate that Pdcd4 prevents ca. 60 to 70% of eIF4A binding to eIF4Gc at a Pdcd4/eIF4A ratio of 1:1, but mutants Pdcd4D253A and Pdcd4D253A,D418A do not. Translation of stem-loop structured mRNA is susceptible to inhibition by wild-type Pdcd4 but not by Pdcd4D253A, Pdcd4D418A, or Pdcd4D235A,D418A. Together, these results indicate that not only binding to eIF4A but also prevention of eIF4A binding to the MA-3 domain of eIF4Gc contributes to the mechanism by which Pdcd4 inhibits translation.
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characterization of programmed cell death 4 in multiple human cancers reveals a novel enhancer of drug sensitivity
Molecular Cancer Therapeutics, 2004Co-Authors: Aaron P Jansen, Corinne E Camalier, Cristi Stark, Nancy H ColburnAbstract:Programmed cell death 4 (Pdcd4), originally identified as an inhibitor of murine cellular transformation, inhibits protein synthesis by directly interacting with eukaryotic Initiation Factor 4A (eIF4A) of the translation Initiation complex. The relevance of Pdcd4 to a broad range of human cancers derived from multiple tissue sites is unknown. Protein expression patterns from the National Cancer Institute drug-screening panel of 60 human cancer cells (NCI60) were analyzed by Western blot methods and revealed frequent reduction of Pdcd4 protein levels in renal-, lung-, and glia-derived tumors. Greater than mean Pdcd4 protein levels correlated with the antitumor activity of geldanamycin and tamoxifen. Stable expression of antisense PDCD4 significantly reduced the sensitivity of MCF-7 breast cancer cells to geldanamycin and to tamoxifen. Sensitivity to geldanamycin significantly increased in UO-31 renal cancer cells expressing sense PDCD4 cDNA. Increased geldanamycin sensitivity was accompanied by enhanced cell cycle arrest and apoptosis. One primary mode of inactivation of Pdcd4 in human cancers appears to involve down-regulated expression, and this down-regulation causes a decreased sensitivity to geldanamycin cytotoxicity. Thus, up-regulating Pdcd4 expression may be promising for geldanamycin-based combination therapy.
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the transformation suppressor pdcd4 is a novel eukaryotic translation Initiation Factor 4A binding protein that inhibits translation
Molecular and Cellular Biology, 2003Co-Authors: Hsinsheng Yang, William C. Merrick, Nahum Sonenberg, Aaron P Jansen, Anton A Komar, Xiaojing Zheng, Sylvain V Costes, Stephen J Lockett, Nancy H ColburnAbstract:Pdcd4 is a novel transformation suppressor that inhibits tumor promoter-induced neoplastic transformation and the activation of AP-1-dependent transcription required for transformation. A yeast two-hybrid analysis revealed that Pdcd4 associates with the eukaryotic translation Initiation Factors eIF4AI and eIF4AII. Immunofluorescent confocal microscopy showed that Pdcd4 colocalizes with eIF4A in the cytoplasm. eIF4A is an ATP-dependent RNA helicase needed to unwind 5' mRNA secondary structure. Recombinant Pdcd4 specifically inhibited the helicase activity of eIF4A and eIF4F. In vivo translation assays showed that Pdcd4 inhibited cap-dependent but not internal ribosome entry site (IRES)-dependent translation. In contrast, Pdcd4(D418A), a mutant inactivated for binding to eIF4A, failed to inhibit cap-dependent or IRES-dependent translation or AP-1 transactivation. Recombinant Pdcd4 prevented eIF4A from binding to the C-terminal region of eIF4G (amino acids 1040 to 1560) but not to the middle region of eIF4G(amino acids 635 to 1039). In addition, both Pdcd4 and Pdcd4(D418A) bound to the middle region of eIF4G. The mechanism by which Pdcd4 inhibits translation thus appears to involve inhibition of eIF4A helicase, interference with eIF4A association-dissociation from eIF4G, and inhibition of eIF4A binding to the C-terminal domain of eIF4G. Pdcd4 binding to eIF4A is linked to its transformation-suppressing activity, as Pdcd4-eIF4A binding and consequent inhibition of translation are required for Pdcd4 transrepression of AP-1.