Academic Journal
Evidence That Eukaryotic Translation Elongation Factor 1A (eEF1A) Binds the Gcn2 Protein C Terminus and Inhibits Gcn2 Activity
| العنوان: | Evidence That Eukaryotic Translation Elongation Factor 1A (eEF1A) Binds the Gcn2 Protein C Terminus and Inhibits Gcn2 Activity |
|---|---|
| المؤلفون: | Visweswaraiah, Jyothsna, Lageix, Sebastien, Castilho, Beatriz Amaral de UNIFESP, Izotova, Lara, Kinzy, Terri Goss, Hinnebusch, Alan G., Sattlegger, Evelyn |
| المساهمون: | Massey Univ, NICHD, Universidade Federal de São Paulo (UNIFESP), Univ Med & Dent New Jersey |
| بيانات النشر: | Amer Soc Biochemistry Molecular Biology Inc |
| سنة النشر: | 2011 |
| المجموعة: | Universidade Federal de São Paulo (UNIFESP): Repositório Institucional |
| الوصف: | The eukaryotic elongation factor 1A (eEF1A) delivers aminoacyl-tRNAs to the ribosomal A-site during protein synthesis. To ensure a continuous supply of amino acids, cells harbor the kinase Gcn2 and its effector protein Gcn1. the ultimate signal for amino acid shortage is uncharged tRNAs. We have proposed a model for sensing starvation, in which Gcn1 and Gcn2 are tethered to the ribosome, and Gcn1 is directly involved in delivering uncharged tRNAs from the A-site to Gcn2 for its subsequent activation. Gcn1 and Gcn2 are large proteins, and these proteins as well as eEF1A access the A-site, leading us to investigate whether there is a functional or physical link between these proteins. Using Saccharomyces cerevisiae cells expressing His(6)-eEF1A and affinity purification, we found that eEF1A co-eluted with Gcn2. Furthermore, Gcn2 co-immunoprecipitated with eEF1A, suggesting that they reside in the same complex. the purified GST-tagged Gcn2 C-terminal domain (CTD) was sufficient for precipitating eEF1A from whole cell extracts generated from gcn2 Delta cells, independently of ribosomes. Purified GST-Gcn2-CTD and purified His(6)-eEF1A interacted with each other, and this was largely independent of the Lys residues in Gcn2-CTD known to be required for tRNA binding and ribosome association. Interestingly, Gcn2-eEF1A interaction was diminished in amino acid-starved cells and by uncharged tRNAs in vitro, suggesting that eEF1A functions as a Gcn2 inhibitor. Consistent with this possibility, purified eEF1A reduced the ability of Gcn2 to phosphorylate its substrate, eIF2 alpha, but did not diminish Gcn2 autophosphorylation. These findings implicate eEF1A in the intricate regulation of Gcn2 and amino acid homeostasis. ; National Institutes of Health ; Fundacao de Apoio a Pesquisa no Estado de São Paulo ; Marsden Fund Council ; Massey University ; Royal Society of New Zealand ; Massey Univ, Inst Nat Sci, N Shore Mail Ctr, Albany 0745, New Zealand ; NICHD, Lab Gene Regulat & Dev, NIH, Bethesda, MD 20892 USA ; Universidade ... |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | 36568-36579 |
| اللغة: | English |
| تدمد: | 0021-9258 |
| Relation: | Journal of Biological Chemistry; http://dx.doi.org/10.1074/jbc.M111.248898; Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 286, n. 42, p. 36568-36579, 2011.; http://repositorio.unifesp.br/handle/11600/34151; WOS:000296538300041 |
| DOI: | 10.1074/jbc.M111.248898 |
| الاتاحة: | http://repositorio.unifesp.br/handle/11600/34151 https://doi.org/10.1074/jbc.M111.248898 |
| Rights: | Acesso aberto |
| رقم الانضمام: | edsbas.CEC08FDE |
| قاعدة البيانات: | BASE |
Full Text Finder | تدمد: | 00219258 |
|---|---|
| DOI: | 10.1074/jbc.M111.248898 |