Structural Basis for the Enhancement of EIF4A Helicase Activity by EIF4G

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2005 Sep 17

PMID 16166382

Citations 78

Authors

Monika Oberer

Assen Marintchev

Gerhard Wagner

Affiliations

Soon will be listed here.

Abstract

The eukaryotic translation initiation factors 4A (eIF4A) and 4G (eIF4G) are crucial for the assembly of the translationally active ribosome. Together with eIF4E, they form the eIF4F complex, which recruits the 40S subunit to the 5' cap of mRNA. The two-domain RNA helicase eIF4A is a very weak helicase by itself, but the activity is enhanced upon interaction with the scaffolding protein eIF4G. Here we show that, albeit both eIF4A domains play a role in binding the middle domain of eIF4G (eIF4G-m, amino acids 745-1003), the main interaction surface is located on the C-terminal domain. We use NMR spectroscopy to define the binding site and find that the contact surface is adjacent to the RNA-, ATP-, and eIF4A-NTD-interacting regions. Mutations of interface residues abrogated binding, confirmed the interface, and showed that the N-terminal end of eIF4G-m interacts with the C-terminal domain of eIF4A. The data suggest that eIF4G-m forms a soft clamp to stabilize the closed interdomain orientation of eIF4A. This model can explain the cooperativity between all binding partners of eIF4A (eIF4G, RNA, ATP) and stimulation of eIF4A activity in the eIF4F complex.

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References

Rogers Jr G, Richter N, Lima W, Merrick W . Modulation of the helicase activity of eIF4A by eIF4B, eIF4H, and eIF4F. J Biol Chem. 2001; 276(33):30914-22. DOI: 10.1074/jbc.M100157200. View

Cornilescu G, Delaglio F, Bax A . Protein backbone angle restraints from searching a database for chemical shift and sequence homology. J Biomol NMR. 1999; 13(3):289-302. DOI: 10.1023/a:1008392405740. View

Rogers Jr G, Komar A, Merrick W . eIF4A: the godfather of the DEAD box helicases. Prog Nucleic Acid Res Mol Biol. 2002; 72:307-31. DOI: 10.1016/s0079-6603(02)72073-4. View

Ali I, McKendrick L, Morley S, Jackson R . Truncated initiation factor eIF4G lacking an eIF4E binding site can support capped mRNA translation. EMBO J. 2001; 20(15):4233-42. PMC: 149147. DOI: 10.1093/emboj/20.15.4233. View

Linder P, Lasko P, Ashburner M, Leroy P, Nielsen P, Nishi K . Birth of the D-E-A-D box. Nature. 1989; 337(6203):121-2. DOI: 10.1038/337121a0. View

Bartels C, Xia T, Billeter M, Guntert P, Wuthrich K . The program XEASY for computer-supported NMR spectral analysis of biological macromolecules. J Biomol NMR. 2012; 6(1):1-10. DOI: 10.1007/BF00417486. View

Kim J, Morgenstern K, Griffith J, Dwyer M, Thomson J, Murcko M . Hepatitis C virus NS3 RNA helicase domain with a bound oligonucleotide: the crystal structure provides insights into the mode of unwinding. Structure. 1998; 6(1):89-100. DOI: 10.1016/s0969-2126(98)00010-0. View

Delaglio F, Grzesiek S, Vuister G, Zhu G, Pfeifer J, Bax A . NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR. 1995; 6(3):277-93. DOI: 10.1007/BF00197809. View

Yang H, Jansen A, Komar A, Zheng X, Merrick W, Costes S . The transformation suppressor Pdcd4 is a novel eukaryotic translation initiation factor 4A binding protein that inhibits translation. Mol Cell Biol. 2002; 23(1):26-37. PMC: 140682. DOI: 10.1128/MCB.23.1.26-37.2003. View

10.

Gross J, Moerke N, von der Haar T, Lugovskoy A, Sachs A, McCarthy J . Ribosome loading onto the mRNA cap is driven by conformational coupling between eIF4G and eIF4E. Cell. 2003; 115(6):739-50. DOI: 10.1016/s0092-8674(03)00975-9. View

11.

Korneeva N, First E, Benoit C, Rhoads R . Interaction between the NH2-terminal domain of eIF4A and the central domain of eIF4G modulates RNA-stimulated ATPase activity. J Biol Chem. 2004; 280(3):1872-81. DOI: 10.1074/jbc.M406168200. View

12.

Marintchev A, Wagner G . Translation initiation: structures, mechanisms and evolution. Q Rev Biophys. 2005; 37(3-4):197-284. DOI: 10.1017/S0033583505004026. View

13.

Caruthers J, Johnson E, McKay D . Crystal structure of yeast initiation factor 4A, a DEAD-box RNA helicase. Proc Natl Acad Sci U S A. 2000; 97(24):13080-5. PMC: 27181. DOI: 10.1073/pnas.97.24.13080. View

14.

Story R, Weber I, Steitz T . The structure of the E. coli recA protein monomer and polymer. Nature. 1992; 355(6358):318-25. DOI: 10.1038/355318a0. View

15.

Marcotrigiano J, Lomakin I, Sonenberg N, Pestova T, Hellen C, Burley S . A conserved HEAT domain within eIF4G directs assembly of the translation initiation machinery. Mol Cell. 2001; 7(1):193-203. DOI: 10.1016/s1097-2765(01)00167-8. View

16.

Tanner N, Cordin O, Banroques J, Doere M, Linder P . The Q motif: a newly identified motif in DEAD box helicases may regulate ATP binding and hydrolysis. Mol Cell. 2003; 11(1):127-38. DOI: 10.1016/s1097-2765(03)00006-6. View

17.

Svitkin Y, Pause A, Haghighat A, Pyronnet S, Witherell G, Belsham G . The requirement for eukaryotic initiation factor 4A (elF4A) in translation is in direct proportion to the degree of mRNA 5' secondary structure. RNA. 2001; 7(3):382-94. PMC: 1370095. DOI: 10.1017/s135583820100108x. View

18.

Imataka H, Sonenberg N . Human eukaryotic translation initiation factor 4G (eIF4G) possesses two separate and independent binding sites for eIF4A. Mol Cell Biol. 1997; 17(12):6940-7. PMC: 232551. DOI: 10.1128/MCB.17.12.6940. View

19.

Delagoutte E, von Hippel P . Helicase mechanisms and the coupling of helicases within macromolecular machines. Part I: Structures and properties of isolated helicases. Q Rev Biophys. 2003; 35(4):431-78. DOI: 10.1017/s0033583502003852. View

20.

Zakowicz H, Yang H, Stark C, Wlodawer A, LaRonde-LeBlanc N, Colburn N . Mutational analysis of the DEAD-box RNA helicase eIF4AII characterizes its interaction with transformation suppressor Pdcd4 and eIF4GI. RNA. 2005; 11(3):261-74. PMC: 1370716. DOI: 10.1261/rna.7191905. View