Home LiteratureArticle Details
PMID: 2304461 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Bidirectional RNA helicase activity of eucaryotic translation initiation factors 4A and 4F.

Molecular and cellular biology ·Vol. 10 ·No. 3 ·1990-03-00 ·Pages 1134-44

Rozen F, Edery I, Meerovitch K, Dever TE, Merrick WC, Sonenberg N

Abstract

The mechanism of ribosome binding to eucaryotic mRNAs is not well understood, but it requires the participation of eucaryotic initiation factors eIF-4A, eIF-4B, and eIF-4F and the hydrolysis of ATP. Evidence has accumulated in support of a model in which these initiation factors function to unwind the 5'-proximal secondary structure in mRNA to facilitate ribosome binding. To obtain direct evidence for initiation factor-mediated RNA unwinding, we developed a simple assay to determine RNA helicase activity, and we show that eIF-4A or eIF-4F, in combination with eIF-4B, exhibits helicase activity. A striking and unprecedented feature of this activity is that it functions in a bidirectional manner. Thus, unwinding can occur either in the 5'-to-3' or 3'-to-5' direction. Unwinding in the 5'-to-3' direction by eIF-4F (the cap-binding protein complex), in conjunction with eIF-4B, was stimulated by the presence of the RNA 5' cap structure, whereas unwinding in the 3'-to-5' direction was completely cap independent. These results are discussed with respect to cap-dependent versus cap-independent mechanisms of ribosome binding to eucaryotic mRNAs.

MeSH Terms
Animals Base Sequence Cell-Free System Eukaryotic Initiation Factor-4A Eukaryotic Initiation Factor-4F In Vitro Techniques Molecular Sequence Data Peptide Chain Initiation, Translational Peptide Initiation Factors/metabolism RNA Caps/metabolism RNA Nucleotidyltransferases/metabolism RNA, Double-Stranded/metabolism Rabbits Structure-Activity Relationship Substrate Specificity
Chemicals
Eukaryotic Initiation Factor-4F Peptide Initiation Factors RNA Caps RNA, Double-Stranded Eukaryotic Initiation Factor-4A RNA Nucleotidyltransferases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Rozen F
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Edery I
Meerovitch K
Dever T E
Merrick W C
Sonenberg N
References (63)
63 references, click to expand
  1. Introduction of homologous DNA sequences into mammalian cells induces mutations in the cognate gene.
    Nature. 1986 Nov 6-12;324(6092):34-8 PMID: 3785372
  2. Probing the function of the eucaryotic 5' cap structure by using a monoclonal antibody directed against cap-binding proteins.
    Cell. 1981 Dec;27(3 Pt 2):563-72 PMID: 6101207
  3. Interaction of Escherichia coli dnaB and dnaC(D) gene products in vitro.
    Proc Natl Acad Sci U S A. 1975 Mar;72(3):921-5 PMID: 1093174
  4. 5'-terminal structure of poliovirus polyribosomal RNA is pUp.
    Proc Natl Acad Sci U S A. 1976 Feb;73(2):327-30 PMID: 174102
  5. The 5' end of poliovirus mRNA is not capped with m7G(5')ppp(5')Np.
    Proc Natl Acad Sci U S A. 1976 Feb;73(2):375-80 PMID: 174106
  6. Absence of 5' terminal capping in encephalomyocarditis virus RNA.
    Nucleic Acids Res. 1976 Oct;3(10):2771-87 PMID: 186763
  7. Capping of eucaryotic mRNAs.
    Cell. 1976 Dec;9(4 PT 2):645-53 PMID: 1017010
  8. Nonspecific effect of m7GMP on protein-RNA interactions.
    J Biol Chem. 1978 Oct 10;253(19):6630-2 PMID: 211125
  9. A polypeptide in eukaryotic initiation factors that crosslinks specifically to the 5'-terminal cap in mRNA.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4843-7 PMID: 217002
  10. Purification of a factor that restores translation of vesicular stomatitis virus mRNA in extracts from poliovirus-infected HeLa cells.
    Proc Natl Acad Sci U S A. 1980 Feb;77(2):770-4 PMID: 6244584
  11. 5'-terminal cap structure in eucaryotic messenger ribonucleic acids.
    Microbiol Rev. 1980 Jun;44(2):175-205 PMID: 6247631
  12. The regulation of initiation of mammalian protein synthesis.
    Prog Nucleic Acid Res Mol Biol. 1981;25:127-85 PMID: 6164076
  13. ATP/Mg++-dependent cross-linking of cap binding proteins to the 5' end of eukaryotic mRNA.
    Nucleic Acids Res. 1981 Apr 10;9(7):1643-56 PMID: 7232223
  14. Two forms of purified m7G-cap binding protein with different effects on capped mRNA translation in extracts of uninfected and poliovirus-infected HeLa cells.
    J Biol Chem. 1981 Aug 10;256(15):7691-4 PMID: 6267020
  15. Proteins controlling the helical structure of DNA.
    Annu Rev Biochem. 1981;50:233-60 PMID: 6267987
  16. Regulated phosphorylation and low abundance of HeLa cell initiation factor eIF-4F suggest a role in translational control. Heat shock effects on eIF-4F.
    J Biol Chem. 1987 Jan 5;262(1):380-8 PMID: 3793730
  17. The ATP-dependent interaction of eukaryotic initiation factors with mRNA.
    J Biol Chem. 1987 Mar 15;262(8):3826-32 PMID: 2950099
  18. Transcription termination factor rho is an RNA-DNA helicase.
    Cell. 1987 Mar 27;48(6):945-52 PMID: 3030561
  19. Unusual requirements for optimum translation of polio viral RNA in vitro.
    J Biol Chem. 1983 Jun 10;258(11):7195-9 PMID: 6304083
  20. New initiation factor activity required for globin mRNA translation.
    J Biol Chem. 1983 May 10;258(9):5804-10 PMID: 6853548
  21. Involvement of eukaryotic initiation factor 4A in the cap recognition process.
    J Biol Chem. 1983 Sep 25;258(18):11398-403 PMID: 6604056
  22. 5'-Conformation of capped alfalfa mosaic virus ribonucleic acid 4 may reflect its independence of the cap structure or of cap-binding protein for efficient translation.
    Biochemistry. 1983 Oct 25;22(22):5157-64 PMID: 6317016
  23. Messenger RNA splicing in vitro: an excised intervening sequence and a potential intermediate.
    Cell. 1984 Jun;37(2):415-27 PMID: 6722880
  24. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold.
    EMBO J. 1982;1(8):945-51 PMID: 6329717
  25. RNA-stimulated ATPase activity of eukaryotic initiation factors.
    J Biol Chem. 1984 Jul 10;259(13):8648-54 PMID: 6145716
  26. Demonstration in vitro that eucaryotic initiation factor 3 is active but that a cap-binding protein complex is inactive in poliovirus-infected HeLa cells.
    J Virol. 1984 Sep;51(3):832-7 PMID: 6088805
  27. Functional characterization of eukaryotic mRNA cap binding protein complex: effects on translation of capped and naturally uncapped RNAs.
    Biochemistry. 1984 May 22;23(11):2456-62 PMID: 6089873
  28. Recognition of cap structure in splicing in vitro of mRNA precursors.
    Cell. 1984 Oct;38(3):731-6 PMID: 6567484
  29. mRNA cap binding proteins: essential factors for initiating translation.
    Cell. 1985 Feb;40(2):223-4 PMID: 3967293
  30. Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency.
    Cell. 1985 Mar;40(3):515-26 PMID: 2982496
  31. ATP-dependent unwinding of messenger RNA structure by eukaryotic initiation factors.
    J Biol Chem. 1985 Jun 25;260(12):7651-8 PMID: 3838990
  32. Eukaryotic protein synthesis.
    Annu Rev Biochem. 1985;54:1109-49 PMID: 3896117
  33. Photoaffinity labeling of the cap-binding protein complex with ATP/dATP. Differential labeling of free eukaryotic initiation factor 4A and the eukaryotic initiation factor 4A component of the cap-binding protein complex with [alpha-32P]ATP/dATP.
    J Biol Chem. 1985 Nov 5;260(25):13831-7 PMID: 4055759
  34. Cloning of eukaryotic protein synthesis initiation factor genes: isolation and characterization of cDNA clones encoding factor eIF-4A.
    Nucleic Acids Res. 1985 Oct 11;13(19):6867-80 PMID: 3840589
  35. Internal initiation of translation on the vesicular stomatitis virus phosphoprotein mRNA yields a second protein.
    J Virol. 1986 Jun;58(3):797-804 PMID: 3009888
  36. Initiation of protein synthesis in mammalian cells.
    Biochem J. 1986 May 1;235(3):625-37 PMID: 3530248
  37. DNA helicase activity of SV40 large tumor antigen.
    EMBO J. 1986 Aug;5(8):1939-44 PMID: 3019672
  38. Effect of upstream reading frames on translation efficiency in simian virus 40 recombinants.
    Mol Cell Biol. 1986 Jul;6(7):2704-11 PMID: 3023946
  39. GTP-binding domain: three consensus sequence elements with distinct spacing.
    Proc Natl Acad Sci U S A. 1987 Apr;84(7):1814-8 PMID: 3104905
  40. A fluorescence study of the binding of eucaryotic initiation factors to messenger RNA and messenger RNA analogues.
    Biochemistry. 1987 Mar 24;26(6):1551-6 PMID: 3593677
  41. Purification and characterization of a new DNA-dependent ATPase with helicase activity from Escherichia coli.
    J Biol Chem. 1987 Nov 5;262(31):15269-76 PMID: 2822720
  42. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates.
    Nucleic Acids Res. 1987 Nov 11;15(21):8783-98 PMID: 3684574
  43. The unwinding of duplex regions in DNA by the simian virus 40 large tumor antigen-associated DNA helicase activity.
    J Biol Chem. 1988 Jan 5;263(1):383-92 PMID: 2826443
  44. Nuclear protein with sequence homology to translation initiation factor eIF-4A.
    Nature. 1988 Apr 21;332(6166):736-8 PMID: 2451786
  45. Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2523-7 PMID: 2833742
  46. Biochemical evidence supporting a mechanism for cap-independent and internal initiation of eukaryotic mRNA.
    J Biol Chem. 1988 May 5;263(13):6016-9 PMID: 2966150
  47. Discriminatory interaction of purified eukaryotic initiation factors 4F plus 4A with the 5' ends of reovirus messenger RNAs.
    J Biol Chem. 1988 May 25;263(15):7266-76 PMID: 3366779
  48. A segment of the 5' nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation.
    J Virol. 1988 Aug;62(8):2636-43 PMID: 2839690
  49. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA.
    Nature. 1988 Jul 28;334(6180):320-5 PMID: 2839775
  50. Leader length and secondary structure modulate mRNA function under conditions of stress.
    Mol Cell Biol. 1988 Jul;8(7):2737-44 PMID: 3405216
  51. Mitochondrial splicing requires a protein from a novel helicase family.
    Nature. 1989 Jan 5;337(6202):84-7 PMID: 2535893
  52. Birth of the D-E-A-D box.
    Nature. 1989 Jan 12;337(6203):121-2 PMID: 2563148
  53. The dnaB-dnaC replication protein complex of Escherichia coli. II. Role of the complex in mobilizing dnaB functions.
    J Biol Chem. 1989 Feb 15;264(5):2469-75 PMID: 2536713
  54. Cap-binding proteins of eukaryotic messenger RNA: functions in initiation and control of translation.
    Prog Nucleic Acid Res Mol Biol. 1988;35:173-207 PMID: 3065823
  55. An essential yeast protein, encoded by duplicated genes TIF1 and TIF2 and homologous to the mammalian translation initiation factor eIF-4A, can suppress a mitochondrial missense mutation.
    Proc Natl Acad Sci U S A. 1989 Apr;86(7):2286-90 PMID: 2648398
  56. Cap recognition and the entry of mRNA into the protein synthesis initiation cycle.
    Trends Biochem Sci. 1988 Feb;13(2):52-6 PMID: 3238751
  57. Identification and purification of a protein that stimulates the helicase activity of the Escherichia coli Rep protein.
    J Biol Chem. 1989 Apr 15;264(11):6119-26 PMID: 2522929
  58. RNA helicase activity associated with the human p68 protein.
    Nature. 1989 Jun 15;339(6225):562-4 PMID: 2471939
  59. RNA unwinding activity of SV40 large T antigen.
    Cell. 1989 Jun 16;57(6):955-63 PMID: 2472217
  60. The Escherichia coli primosome can translocate actively in either direction along a DNA strand.
    J Biol Chem. 1989 Aug 25;264(24):14531-42 PMID: 2547799
  61. Dissociation of double-stranded polynucleotide helical structures by eukaryotic initiation factors, as revealed by a novel assay.
    Biochemistry. 1989 May 30;28(11):4729-34 PMID: 2548591
  62. A lysine substitution in the ATP-binding site of eucaryotic initiation factor 4A abrogates nucleotide-binding activity.
    Mol Cell Biol. 1989 Sep;9(9):4061-3 PMID: 2506440
  63. Tobacco mosaic virus ribonucleic acid-dependent amino acid incorporation in a wheat embryo system in vitro. Analysis of the rate-limiting reaction.
    J Biol Chem. 1970 Mar 10;245(5):955-61 PMID: 5417266
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.3.1134-1144.1990
Published
1990-03-00
Pages
1134-44
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360981
Subset
IM
Grants
NIADDK NIH HHS · AM07319 · United States
NIGMS NIH HHS · GM26796 · United States
External Links
PubMed source DOI: 10.1128/mcb.10.3.1134-1144.1990 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2