@article{c62fb4e5d8b543f8b6a978404bc5b307,
title = "The C-Terminal Domain of Eukaryotic Initiation Factor 5 Promotes Start Codon Recognition by Its Dynamic Interplay with eIF1 and eIF2β",
abstract = "Recognition of the proper start codon on mRNAs is essential for protein synthesis, which requires scanning and involves eukaryotic initiation factors (eIFs) eIF1, eIF1A, eIF2, and eIF5. The carboxyl terminal domain (CTD) of eIF5 stimulates 43S preinitiation complex (PIC) assembly; however, its precise role in scanning and start codon selection has remained unknown. Using nuclear magnetic resonance (NMR) spectroscopy, we identified the binding sites of eIF1 and eIF2β on eIF5-CTD and found that they partially overlapped. Mutating select eIF5 residues in the common interface specifically disrupts interaction with both factors. Genetic and biochemical evidence indicates that these eIF5-CTD mutations impair start codon recognition and impede eIF1 release from the PIC by abrogating eIF5-CTD binding to eIF2β. This study provides mechanistic insight into the role of eIF5-CTD@s dynamic interplay with eIF1 and eIF2β in switching PICs from an open to a closed state at start codons.",
author = "Luna, {Rafael E.} and Haribabu Arthanari and Hiroyuki Hiraishi and Jagpreet Nanda and Pilar Martin-Marcos and Markus, {Michelle A.} and Barak Akabayov and Milbradt, {Alexander G.} and Luna, {Lunet E.} and Seo, {Hee Chan} and Hyberts, {Sven G.} and Amr Fahmy and Mikhail Reibarkh and David Miles and Hagner, {Patrick R.} and O'Day, {Elizabeth M.} and Tingfang Yi and Assen Marintchev and Hinnebusch, {Alan G.} and Lorsch, {Jon R.} and Katsura Asano and Gerhard Wagner",
note = "Funding Information: This work was supported by NIH Grants CA68262 and GM47467 to G.W., NRSA-GM79970 to R.E.L., NIDDK-K01-DK085198 to H.A., G.W.'s GM47467 for L.E.L., GM64781 to K.A., NHBLI-T32HL07623-25 for P.R.H., and by the Intramural Research Program of the NIH (to P.M.-M. and A.G.H.). We thank T. Pestova for generously providing us with eIF1 mutant constructs. We would like to also thank L. Yang and M. Allier (Beamline X-9 NSLS, BNL) at Brookhaven National Laboratory. Use of the National Synchrotron Light Source, Brookhaven National Laboratory, was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract No. DE-AC02-98CH10886. We thank E. Bedoya for purifying proteins and V. D'Souza and M. Durney for help with ITC data. ",
year = "2012",
month = jun,
day = "28",
doi = "10.1016/j.celrep.2012.04.007",
language = "English (US)",
volume = "1",
pages = "689--702",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "6",
}