2014
DOI: 10.1128/mcb.00663-14
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Functional and Biochemical Characterization of Human Eukaryotic Translation Initiation Factor 3 in Living Cells

Abstract: The main role of the translation initiation factor 3 (eIF3) is to orchestrate formation of 43S-48S preinitiation complexes (PICs). Until now, most of our knowledge on eIF3 functional contribution to regulation of gene expression comes from yeast studies. Hence, here we developed several novel in vivo assays to monitor the integrity of the 13-subunit human eIF3 complex, defects in assembly of 43S PICs, efficiency of mRNA recruitment, and postassembly events such as AUG recognition. We knocked down expression of… Show more

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Cited by 75 publications
(151 citation statements)
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“…58,59,63,99 Furthermore, eIF3A, the largest and conserved eIF3 subunit, may control the assembly and stability of eIF3 as well as the formation of different sub-complexes with additional functions within the mammalian cells. 64,100 However, differently from the Drosophila homolog, ABCE1 silencing in our system did not reduce the levels of eIF3A and eIF3H (Fig. 6A), as previously demonstrated also for several initiation factors in yeast.…”
Section: Discussionsupporting
confidence: 84%
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“…58,59,63,99 Furthermore, eIF3A, the largest and conserved eIF3 subunit, may control the assembly and stability of eIF3 as well as the formation of different sub-complexes with additional functions within the mammalian cells. 64,100 However, differently from the Drosophila homolog, ABCE1 silencing in our system did not reduce the levels of eIF3A and eIF3H (Fig. 6A), as previously demonstrated also for several initiation factors in yeast.…”
Section: Discussionsupporting
confidence: 84%
“…62 For further studies on human ABCE1 interacting partners, we selected the eIF3A and eIF3C subunits, which belong to the functional core of mammalian eIF3 complex and play a critical role in its integrity. 63,64 Co-immunoprecipitations with anti-ABCE1 antibody were performed on HEK293 whole cell lysates, and the precipitated proteins were analyzed by Western blot. Both eIF3A and eIF3C were detected at rather low intensity in several experiments (Fig.…”
Section: Abce1 Downregulation Impairs Cell Proliferation and Cell Cycmentioning
confidence: 99%
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“…As a consequence, eIF3 is thought to stabilize mRNA binding to the ribosome. Whether human eIF3 directly affects downstream reactions in initiation is not yet firmly established, although knockdown of the 3c subunit in HeLa cells results in a relaxed stringency of start codon selection [25]. eIF3 is expected to dissociate following formation of the 80S initiation complex, although this has not been studied carefully.…”
Section: The Role Of Human Eif3 In Protein Synthesismentioning
confidence: 99%
“…The importance of 3j in mRNA recruitment is challenged, however, by the finding that a 5-fold siRNA knockdown of 3j levels in HeLa or HEK293 cells causes only a minor reduction in protein synthesis [25]. mRNA protruding from the 40S mRNA exit site interacts with 3a and 3d, likely by extending into the solvent side of the ribosomal subunit were the bulk of the eIF3 is located [26].…”
Section: The Role Of Human Eif3 In Protein Synthesismentioning
confidence: 99%