Eif3c Antibody A
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Structural Characterization of the Human Eukaryotic Initiation Factor 3 Protein Complex by Mass Spectrometry*□S
Supplemental Material can be found at: http://www.mcponline.org/cgi/content/full/M600399-MCP200 /DC1 Research Structural Characterization of the Human Eukaryotic Initiation Factor 3 Protein Complex by Mass Spectrometry*□S Eugen Damoc‡, Christopher S. Fraser§, Min Zhou¶, Hortense Videler¶, Greg L. Mayeurʈ, John W. B. Hersheyʈ, Jennifer A. Doudna§, Carol V. Robinson¶**, and Julie A. Leary‡ ‡‡ Protein synthesis in mammalian cells requires initiation The initiation phase of eukaryotic protein synthesis involves factor eIF3, an ϳ800-kDa protein complex that plays a formation of an 80 S ribosomal complex containing the initi- Downloaded from central role in binding of initiator methionyl-tRNA and ator methionyl-tRNAi bound to the initiation codon in the mRNA to the 40 S ribosomal subunit to form the 48 S mRNA. This is a multistep process promoted by proteins initiation complex. The eIF3 complex also prevents pre- called eukaryotic initiation factors (eIFs).1 Currently at least 12 mature association of the 40 and 60 S ribosomal subunits eIFs, composed of at least 29 distinct subunits, have been and interacts with other initiation factors involved in start identified (1). Mammalian eIF3, the largest initiation factor, is a codon selection. The molecular mechanisms by which multisubunit complex with an apparent molecular mass of www.mcponline.org eIF3 exerts these functions are poorly understood. Since ϳ800 kDa. This protein complex plays an essential role in its initial characterization in the 1970s, the exact size, translation by binding directly to the 40 S ribosomal subunit composition, and post-translational modifications of and promoting formation of the 43 S preinitiation complex ⅐ ⅐ mammalian eIF3 have not been rigorously determined. -
A Chemical-Genetic Screen for Identifying Substrates of the Er Kinase Perk
University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2014 A Chemical-Genetic Screen for Identifying Substrates of the Er Kinase Perk Nancy L. Maas University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Biology Commons, Cell Biology Commons, and the Molecular Biology Commons Recommended Citation Maas, Nancy L., "A Chemical-Genetic Screen for Identifying Substrates of the Er Kinase Perk" (2014). Publicly Accessible Penn Dissertations. 1354. https://repository.upenn.edu/edissertations/1354 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/1354 For more information, please contact [email protected]. A Chemical-Genetic Screen for Identifying Substrates of the Er Kinase Perk Abstract Cells constantly encounter changing environments that challenge the ability to adapt and survive. Signal transduction networks enable cells to appropriately sense and respond to these changes, and are often mediated through the activity of protein kinases. Protein kinases are a class of enzyme responsible for regulating a broad spectrum of cellular functions by transferring phosphate groups from ATP to substrate proteins, thereby altering substrate activity and function. PERK is a resident kinase of the endoplasmic reticulum, and is responsible for sensing perturbations in the protein folding capacity of the ER. When the influx of unfolded, nascent proteins exceeds the folding capacity of the ER, PERK initiates a cascade of signaling events that enable cell adaptation and ER stress resolution. These signaling pathways are not only essential for the survival of normal cells undergoing ER stress, but are also co-opted by tumor cells in order to survive the oxygen and nutrient-restricted conditions of the tumor microenvironment. -
CHARACTERIZING the INTERACTION BETWEEN PDCD4 and Eif3 with RESPECT to TRANSLATION REGULATION
CHARACTERIZING THE INTERACTION BETWEEN PDCD4 AND eIF3 WITH RESPECT TO TRANSLATION REGULATION DIVYA SHARMA KHANDIGA Master of Science, Bangalore University, India 2009 A Thesis/Project Submitted to the School of Graduate Studies of the University of Lethbridge in Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE Department of Chemistry and Biochemistry University of Lethbridge LETHBRIDGE, ALBERTA, CANADA © Divya Sharma Khandiga, 2017 CHARACTERIZING THE INTERACTION BETWEEN PDCD4 AND eIF3 WITH RESPECT TO TRANSLATION REGULATION DIVYA SHARMA KHANDIGA Date of Defense: December 12, 2017 Dr. N. Thakor Assistant Professor Ph.D. Thesis Supervisor Dr. M. Roussel Professor Ph.D. Thesis Co-supervisor Dr. U. Kothe Associate Professor Ph.D. Thesis Examination Committee Member Dr. R. Golsteyn Associate Professor Ph.D. Thesis Examination Committee Member Dr. R. Fahlman Professor Ph.D. External Examiner University of Alberta Edmonton, Alberta Dr. M. Gerken Professor Ph.D. Chair, Thesis Examination Committee Dedication To my beloved family and friends, My inspiration, my parents Subraya Sharma and Kamala Sharma My dearly loved husband Samarth, sister Dr. Lakshmi and brother-in-law Dr. Pradeep My cute little niece Mithali and nephew Aathreya My adorable brother Dr. Ganesh, sister Dr. Sharadha, Silly Vidya and little angels My loving cousins and in-laws I am grateful to have them in my life, it is their well wishes, teachings, support and love that have enabled me to achieve success and happiness in life. iii Abstract Programmed cell death protein 4 (PDCD4) inhibits IRES-mediated translation of anti- apoptotic proteins such as XIAP. PDCD4 was shown to directly interact with the XIAP IRES element and inhibit translation initiation. -
Genes with 5' Terminal Oligopyrimidine Tracts Preferentially Escape Global Suppression of Translation by the SARS-Cov-2 NSP1 Protein
Downloaded from rnajournal.cshlp.org on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press Genes with 5′ terminal oligopyrimidine tracts preferentially escape global suppression of translation by the SARS-CoV-2 Nsp1 protein Shilpa Raoa, Ian Hoskinsa, Tori Tonna, P. Daniela Garciaa, Hakan Ozadama, Elif Sarinay Cenika, Can Cenika,1 a Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA 1Corresponding author: [email protected] Key words: SARS-CoV-2, Nsp1, MeTAFlow, translation, ribosome profiling, RNA-Seq, 5′ TOP, Ribo-Seq, gene expression 1 Downloaded from rnajournal.cshlp.org on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press Abstract Viruses rely on the host translation machinery to synthesize their own proteins. Consequently, they have evolved varied mechanisms to co-opt host translation for their survival. SARS-CoV-2 relies on a non-structural protein, Nsp1, for shutting down host translation. However, it is currently unknown how viral proteins and host factors critical for viral replication can escape a global shutdown of host translation. Here, using a novel FACS-based assay called MeTAFlow, we report a dose-dependent reduction in both nascent protein synthesis and mRNA abundance in cells expressing Nsp1. We perform RNA-Seq and matched ribosome profiling experiments to identify gene-specific changes both at the mRNA expression and translation level. We discover that a functionally-coherent subset of human genes are preferentially translated in the context of Nsp1 expression. These genes include the translation machinery components, RNA binding proteins, and others important for viral pathogenicity. Importantly, we uncovered a remarkable enrichment of 5′ terminal oligo-pyrimidine (TOP) tracts among preferentially translated genes. -
The Eif3 Complex of Typanosoma Brucei: Composition Conservation Does Not Imply the Conservation of Structural Assembly and Subunits Function
Downloaded from rnajournal.cshlp.org on October 1, 2021 - Published by Cold Spring Harbor Laboratory Press 1 The eIF3 complex of Typanosoma brucei: composition conservation does not imply the 2 conservation of structural assembly and subunits function 3 4 Kunrao Li,1,2 Shuru Zhou,1,2 Qixuan Guo,3 Xin Chen,1,2 Dehua Lai,2,4 Zhaorong Lun,2,4,5 and 5 Xuemin Guo1,2,5 6 7 1Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-Sen University, 8 Guangzhou, China 9 2Key Laboratory of Tropical Disease Control (Sun Yat-Sen University), Ministry of Education, 10 Guangzhou, China 11 3Chengde Nursing Vocational College, Chende, China 12 4Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life Sciences, 13 Sun Yat-Sen University, Guangzhou, China 14 15 5Corresponding author. Email, [email protected]; or [email protected] 16 K.L. and S.Z. contributed equally to this work 17 18 Running head: Characterization of the eIF3 of Trypanosoma brucei 19 Keywords: Translation, Eukaryotic initiation factor 3, Trypanosome 20 1 Downloaded from rnajournal.cshlp.org on October 1, 2021 - Published by Cold Spring Harbor Laboratory Press 21 ABSTRACT 22 The multisubunit eukaryotic initiation factor 3 (eIF3) plays multiple roles in translation, but 23 poorly understood in trypanosomes. The putative subunits eIF3a and eIF3f of Trypanosoma 24 brucei (TbIF3a and TbIF3f) were overexpressed and purified, and 11 subunits were identified, 25 TbIF3a through l minus j, which form a tight complex. Both TbIF3a and TbIF3f are essential for 26 viability of T. brucei. RNAi knockdown of either of them severely reduced total translation and 27 the ratio of polysome/80S peak area. -
WO 2019/079361 Al 25 April 2019 (25.04.2019) W 1P O PCT
(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization I International Bureau (10) International Publication Number (43) International Publication Date WO 2019/079361 Al 25 April 2019 (25.04.2019) W 1P O PCT (51) International Patent Classification: CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, C12Q 1/68 (2018.01) A61P 31/18 (2006.01) DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, C12Q 1/70 (2006.01) HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, (21) International Application Number: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, PCT/US2018/056167 OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, (22) International Filing Date: SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, 16 October 2018 (16. 10.2018) TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (26) Publication Language: English GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, (30) Priority Data: UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, 62/573,025 16 October 2017 (16. 10.2017) US TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, ΓΕ , IS, IT, LT, LU, LV, (71) Applicant: MASSACHUSETTS INSTITUTE OF MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TECHNOLOGY [US/US]; 77 Massachusetts Avenue, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Cambridge, Massachusetts 02139 (US). -
Eukaryotic Translation Initiation Factor 3 Plays Distinct Roles at The
RESEARCH ARTICLE Eukaryotic translation initiation factor 3 plays distinct roles at the mRNA entry and exit channels of the ribosomal preinitiation complex Colin Echeverrı´aAitken1, Petra Beznoskova´ 2, Vladislava Vlcˇkova2, Wen-Ling Chiu3†, Fujun Zhou1, Leosˇ Shivaya Vala´ sˇek2*, Alan G Hinnebusch3*, Jon R Lorsch1* 1Laboratory on the Mechanism and Regulation of Protein Synthesis, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United States; 2Laboratory of Regulation of Gene Expression, Institute of Microbiology ASCR, Prague, Czech Republic; 3Laboratory of Gene Regulation and Development, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United States Abstract Eukaryotic translation initiation factor 3 (eIF3) is a central player in recruitment of the pre-initiation complex (PIC) to mRNA. We probed the effects on mRNA recruitment of a library of S. cerevisiae eIF3 functional variants spanning its 5 essential subunits using an in vitro-reconstituted *For correspondence: valasekl@ system. Mutations throughout eIF3 disrupt its interaction with the PIC and diminish its ability to biomed.cas.cz (LSV); [email protected] (AGH); jon. accelerate recruitment to a native yeast mRNA. Alterations to the eIF3a CTD and eIF3b/i/g . [email protected] (JRL) significantly slow mRNA recruitment, and mutations within eIF3b/i/g destabilize eIF2 GTP Met- tRNAi binding to the PIC. Using model mRNAs lacking contacts with the 40S entry or exit channels, Present address: we uncovered a critical role for eIF3 requiring the eIF3a NTD, in stabilizing mRNA interactions at †PharmaEssentia Corp., Taipei, the exit channel, and an ancillary role at the entry channel requiring residues of the eIF3a CTD. -
Systematically Profiling the Expression of Eif3 Subunits in Glioma Reveals
Chai et al. Cancer Cell Int (2019) 19:155 https://doi.org/10.1186/s12935-019-0867-1 Cancer Cell International PRIMARY RESEARCH Open Access Systematically profling the expression of eIF3 subunits in glioma reveals the expression of eIF3i has prognostic value in IDH-mutant lower grade glioma Rui‑Chao Chai1,4,6†, Ning Wang2†, Yu‑Zhou Chang3, Ke‑Nan Zhang1,6, Jing‑Jun Li1,6, Jun‑Jie Niu5, Fan Wu1,6*, Yu‑Qing Liu1,6* and Yong‑Zhi Wang1,3,4,6* Abstract Background: Abnormal expression of the eukaryotic initiation factor 3 (eIF3) subunits plays critical roles in tumo‑ rigenesis and progression, and also has potential prognostic value in cancers. However, the expression and clinical implications of eIF3 subunits in glioma remain unknown. Methods: Expression data of eIF3 for patients with gliomas were obtained from the Chinese Glioma Genome Atlas (CGGA) (n 272) and The Cancer Genome Atlas (TCGA) (n 595). Cox regression, the receiver operating characteristic (ROC) curves= and Kaplan–Meier analysis were used to study= the prognostic value. Gene oncology (GO) and gene set enrichment analysis (GSEA) were utilized for functional prediction. Results: In both the CGGA and TCGA datasets, the expression levels of eIF3d, eIF3e, eIF3f, eIF3h and eIF3l highly were associated with the IDH mutant status of gliomas. The expression of eIF3b, eIF3i, eIF3k and eIF3m was increased with the tumor grade, and was associated with poorer overall survival [All Hazard ratio (HR) > 1 and P < 0.05]. By contrast, the expression of eIF3a and eIF3l was decreased in higher grade gliomas and was associated with better overall sur‑ vival (Both HR < 1 and P < 0.05). -
PCI Proteins Eif3e and Eif3m Define Distinct Translation Initiation Factor 3 Complexes
PCI Proteins eIF3e and eIF3m Define Distinct Translation Initiation Factor 3 Complexes The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Zhou, Chunshui, Fatih Arslan, Susan Wee, Srinivasan Krishnan, Alexander R. Ivanov, Anna Oliva, Janet Leatherwood, and Dieter A. Wolf. 2005. PCI proteins eIF3e and eIF3m define distinct translation initiation factor 3 complexes. BMC Biology 3:14. Published Version doi:10.1186/1741-7007-3-14 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:4595136 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA BMC Biology BioMed Central Research article Open Access PCI proteins eIF3e and eIF3m define distinct translation initiation factor 3 complexes Chunshui Zhou1,5, Fatih Arslan1, Susan Wee1, Srinivasan Krishnan2, Alexander R Ivanov3, Anna Oliva4, Janet Leatherwood4 and Dieter A Wolf*1,3 Address: 1Department of Genetics and Complex Diseases, Harvard School of Public Health, 665 Huntington Avenue, Boston, Massachusetts, 02115, USA, 2Applied Biosystems Inc., Framingham, Massachusetts, USA, 3Harvard NIEHS Center Proteomics Facility, Harvard School of Public Health, Boston, Massachusetts, USA, 4Department of Molecular Genetics and Microbiology, State University of New York, Stony Brook, New York, USA and 5Department of -
Translation Initiation Factor Eif3h Targets Specific Transcripts To
Translation initiation factor eIF3h targets specific transcripts to polysomes during embryogenesis Avik Choudhuria,b, Umadas Maitraa,1, and Todd Evansb,1 aDepartment of Developmental and Molecular Biology, Albert Einstein College of Medicine of Yeshiva University, Bronx, NY 10461; and bDepartment of Surgery, Weill Cornell Medical College, New York, NY 10065 Edited by Igor B. Dawid, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, and approved April 30, 2013 (received for review February 14, 2013) Eukaryotic translation initiation factor 3 (eIF3) plays a central role eukaryotes. These—eIF3d, eIF3e, eIF3f, eIF3h, eIF3j, eIF3k, in translation initiation and consists of five core (conserved) sub- eIF3l, and eIF3m—were designated “non-core” subunits (4). units present in both budding yeast and higher eukaryotes. Higher In contrast to the budding yeast, the genome of the fission eukaryotic eIF3 contains additional (noncore or nonconserved) yeast Schizosaccharomyces pombe contains structural homologs subunits of poorly defined function, including sub-unit h (eIF3h), of at least five noncore (nonconserved) eIF3 subunits—eIF3d, which in zebrafish is encoded by two distinct genes (eif3ha and eIF3e, eIF3f, eIF3h, and eIF3m. The gene encoding eIF3f is eif3hb). Previously we showed that eif3ha encodes the predominant essential for growth, whereas eIF3d, eIF3e, and eIF3h are dis- isoform during zebrafish embryogenesis and that depletion of this pensable for growth and viability (5–11). However, deleted factor causes defects in the development of the brain and eyes. To strains show specific phenotypes including defects in meiosis/ investigate the molecular mechanism governing this regulation, we sporulation (6, 9, 11). -
Identification of Differentially Expressed Genes and Pathways in Mice Exposed to Mixed Field Neutron/Photon Radiation Constantinos G
Broustas et al. BMC Genomics (2018) 19:504 https://doi.org/10.1186/s12864-018-4884-6 RESEARCHARTICLE Open Access Identification of differentially expressed genes and pathways in mice exposed to mixed field neutron/photon radiation Constantinos G. Broustas1* , Andrew D. Harken2, Guy Garty2 and Sally A. Amundson1 Abstract Background: Radiation exposure due to the detonation of an improvised nuclear device remains a major security concern. Radiation from such a device involves a combination of photons and neutrons. Although photons will make the greater contribution to the total dose, neutrons will certainly have an impact on the severity of the exposure as they have high relative biological effectiveness. Results: We investigated the gene expression signatures in the blood of mice exposed to 3 Gy x-rays, 0.75 Gy of neutrons, or to mixed field photon/neutron with the neutron fraction contributing 5, 15%, or 25% of a total 3 Gy radiation dose. Gene ontology and pathway analysis revealed that genes involved in protein ubiquitination pathways were significantly overrepresented in all radiation doses and qualities. On the other hand, eukaryotic initiation factor 2 (EIF2) signaling pathway was identified as one of the top 10 ranked canonical pathways in neutron, but not pure x-ray, exposures. In addition, the related mTOR and regulation of EIF4/p70S6K pathways were also significantly underrepresented in the exposures with a neutron component, but not in x-ray radiation. The majority of the changed genes in these pathways belonged to the ribosome biogenesis and translation machinery and included several translation initiation factors (e.g. Eif2ak4, Eif3f), as well as 40S and 60S ribosomal subunits (e.g. -
Global Analysis of LARP1 Translation Targets Reveals Tunable and Dynamic Features of 5′ TOP Motifs
Global analysis of LARP1 translation targets reveals tunable and dynamic features of 5′ TOP motifs Lucas Philippea,1, Antonia M. G. van den Elzena,1, Maegan J. Watsona, and Carson C. Thoreena,2 aDepartment of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, CT 06510 Edited by Alan G. Hinnebusch, National Institutes of Health, Bethesda, MD, and approved January 29, 2020 (received for review July 25, 2019) Terminal oligopyrimidine (TOP) motifs are sequences at the 5′ ends recent findings have hinted that the RNA-binding protein La- of mRNAs that link their translation to the mTOR Complex 1 related protein 1 (LARP1) may have a central role (8–10). (mTORC1) nutrient-sensing signaling pathway. They are com- LARP1 is a large protein (150 kDa) with several RNA-binding monly regarded as discrete elements that reside on ∼100 mRNAs domains. Its central region contains a La motif (LaM) domain that mostly encode translation factors. However, the full spectrum that defines the La-related protein (LARP) superfamily, along of TOP sequences and their prevalence throughout the transcrip- with an adjacent RNA recognition motif-like (RRM-L) domain. tome remain unclear, primarily because of uncertainty over the Its C terminus encodes a domain known as the DM15 region. This mechanism that detects them. Here, we globally analyzed trans- domain is unique to LARP1 and its closely related homolog lation targets of La-related protein 1 (LARP1), an RNA-binding pro- LARP1B, and is therefore also known as the LARP1 domain (11). tein and mTORC1 effector that has been shown to repress TOP Several observations suggest that LARP1 directly represses TOP mRNA translation in a few specific cases.