Home LiteratureArticle Details
PMID: 11340157 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Interaction of eukaryotic translation initiation factor 4G with the nuclear cap-binding complex provides a link between nuclear and cytoplasmic functions of the m(7) guanosine cap.

Molecular and cellular biology ·Vol. 21 ·No. 11 ·2001-06-00 ·Pages 3632-41

McKendrick L, Thompson E, Ferreira J, Morley SJ, Lewis JD

Abstract

In eukaryotes the majority of mRNAs have an m(7)G cap that is added cotranscriptionally and that plays an important role in many aspects of mRNA metabolism. The nuclear cap-binding complex (CBC; consisting of CBP20 and CBP80) mediates the stimulatory functions of the cap in pre-mRNA splicing, 3' end formation, and U snRNA export. As little is known about how nuclear CBC mediates the effects of the cap in higher eukaryotes, we have characterized proteins that interact with CBC in HeLa cell nuclear extracts as potential mediators of its function. Using cross-linking and coimmunoprecipitation, we show that eukaryotic translation initiation factor 4G (eIF4G), in addition to its function in the cytoplasm, is a nuclear CBC-interacting protein. We demonstrate that eIF4G interacts with CBC in vitro and that, in addition to its cytoplasmic localization, there is a significant nuclear pool of eIF4G in mammalian cells in vivo. Immunoprecipitation experiments suggest that, in contrast to the cytoplasmic pool, much of the nuclear eIF4G is not associated with eIF4E (translation cap binding protein of eIF4F) but is associated with CBC. While eIF4G stably associates with spliceosomes in vitro and shows close association with spliceosomal snRNPs and splicing factors in vivo, depletion studies show that it does not participate directly in the splicing reaction. Taken together the data indicate that nuclear eIF4G may be recruited to pre-mRNAs via its interaction with CBC and accompanies the mRNA to the cytoplasm, facilitating the switching of CBC for eIF4F. This may provide a mechanism to couple nuclear and cytoplasmic functions of the mRNA cap structure.

MeSH Terms
3T3 Cells Animals Cell Nucleus/metabolism Cross-Linking Reagents Cytoplasm/metabolism Dinucleoside Phosphates/metabolism Eukaryotic Cells Eukaryotic Initiation Factor-4G Fatty Acids, Unsaturated/pharmacology HeLa Cells Humans Mice Peptide Initiation Factors/metabolism RNA Cap-Binding Proteins RNA Caps/metabolism RNA-Binding Proteins/metabolism Ribonucleoproteins, Small Nuclear/metabolism Spliceosomes/metabolism
Chemicals
Cross-Linking Reagents Dinucleoside Phosphates Eukaryotic Initiation Factor-4G Fatty Acids, Unsaturated Peptide Initiation Factors RNA Cap-Binding Proteins RNA Caps RNA-Binding Proteins Ribonucleoproteins, Small Nuclear 7-methyl-diguanosine triphosphate leptomycin B
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
McKendrick L
Department of Biochemistry, School of Biological Sciences, University of Sussex, Falmer, Brighton BN1 9QG, United Kingdom.
Thompson E
Ferreira J
Morley S J
Lewis J D
References (57)
57 references, click to expand
  1. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
    Anal Biochem. 1987 Nov 1;166(2):368-79 PMID: 2449095
  2. From factors to mechanisms: translation and translational control in eukaryotes.
    Curr Opin Genet Dev. 1999 Oct;9(5):515-21 PMID: 10508691
  3. Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus.
    Nature. 1990 Feb 1;343(6257):437-41 PMID: 2137203
  4. Mammalian nuclei contain foci which are highly enriched in components of the pre-mRNA splicing machinery.
    EMBO J. 1991 Jan;10(1):195-206 PMID: 1824936
  5. Nuclear export of different classes of RNA is mediated by specific factors.
    J Cell Biol. 1994 Mar;124(5):627-35 PMID: 7509815
  6. Differential interaction of splicing snRNPs with coiled bodies and interchromatin granules during mitosis and assembly of daughter cell nuclei.
    J Cell Biol. 1994 Jul;126(1):11-23 PMID: 8027171
  7. A nuclear cap binding protein complex involved in pre-mRNA splicing.
    Cell. 1994 Aug 26;78(4):657-68 PMID: 8069914
  8. A conserved epitope on a subset of SR proteins defines a larger family of Pre-mRNA splicing factors.
    J Cell Biol. 1995 May;129(4):899-908 PMID: 7538140
  9. Nuclear eukaryotic initiation factor 4E (eIF4E) colocalizes with splicing factors in speckles.
    J Cell Biol. 2000 Jan 24;148(2):239-47 PMID: 10648556
  10. Promyelocytic leukemia (PML) nuclear bodies are protein structures that do not accumulate RNA.
    J Cell Biol. 2000 Jan 24;148(2):283-92 PMID: 10648561
  11. Changes in integrity and association of eukaryotic protein synthesis initiation factors during apoptosis.
    Eur J Biochem. 2000 Feb;267(4):1083-91 PMID: 10672017
  12. The role of nuclear cap binding protein Cbc1p of yeast in mRNA termination and degradation.
    Mol Cell Biol. 2000 Apr;20(8):2827-38 PMID: 10733586
  13. PHAX, a mediator of U snRNA nuclear export whose activity is regulated by phosphorylation.
    Cell. 2000 Apr 14;101(2):187-98 PMID: 10786834
  14. Pre-mRNA splicing alters mRNP composition: evidence for stable association of proteins at exon-exon junctions.
    Genes Dev. 2000 May 1;14(9):1098-108 PMID: 10809668
  15. Like attracts like: getting RNA processing together in the nucleus.
    Science. 2000 May 26;288(5470):1385-9 PMID: 10827942
  16. A cap-binding protein complex mediating U snRNA export.
    Nature. 1995 Aug 24;376(6542):709-12 PMID: 7651522
  17. A nuclear cap-binding complex binds Balbiani ring pre-mRNA cotranscriptionally and accompanies the ribonucleoprotein particle during nuclear export.
    J Cell Biol. 1996 Apr;133(1):5-14 PMID: 8601613
  18. A nuclear cap-binding complex facilitates association of U1 snRNP with the cap-proximal 5' splice site.
    Genes Dev. 1996 Jul 1;10(13):1683-98 PMID: 8682298
  19. A novel shuttling protein, 4E-T, mediates the nuclear import of the mRNA 5' cap-binding protein, eIF4E.
    EMBO J. 2000 Jun 15;19(12):3142-56 PMID: 10856257
  20. eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation.
    Annu Rev Biochem. 1999;68:913-63 PMID: 10872469
  21. The yeast nuclear cap binding complex can interact with translation factor eIF4G and mediate translation initiation.
    Mol Cell. 2000 Jul;6(1):191-6 PMID: 10949040
  22. Novel eIF4G domain homologues linking mRNA translation with nonsense-mediated mRNA decay.
    Trends Biochem Sci. 2000 Sep;25(9):423-6 PMID: 10973054
  23. The protein Aly links pre-messenger-RNA splicing to nuclear export in metazoans.
    Nature. 2000 Sep 21;407(6802):401-5 PMID: 11014198
  24. Pre-mRNA splicing imprints mRNA in the nucleus with a novel RNA-binding protein that persists in the cytoplasm.
    Mol Cell. 2000 Sep;6(3):673-82 PMID: 11030346
  25. The spliceosome deposits multiple proteins 20-24 nucleotides upstream of mRNA exon-exon junctions.
    EMBO J. 2000 Dec 15;19(24):6860-9 PMID: 11118221
  26. Capping of eucaryotic mRNAs.
    Cell. 1976 Dec;9(4 PT 2):645-53 PMID: 1017010
  27. The addition of 5' cap structures occurs early in hnRNA synthesis and prematurely terminated molecules are capped.
    Cell. 1980 Jan;19(1):69-78 PMID: 7357608
  28. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  29. The structure of mammalian small nuclear ribonucleoproteins. Identification of multiple protein components reactive with anti-(U1)ribonucleoprotein and anti-Sm autoantibodies.
    J Biol Chem. 1984 May 10;259(9):5907-14 PMID: 6232278
  30. Identification of nuclear cap specific proteins in HeLa cells.
    Nucleic Acids Res. 1987 Aug 25;15(16):6489-500 PMID: 3627995
  31. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates.
    Nucleic Acids Res. 1987 Nov 11;15(21):8783-98 PMID: 3684574
  32. A yeast cap binding protein complex (yCBC) acts at an early step in pre-mRNA splicing.
    Nucleic Acids Res. 1996 Sep 1;24(17):3332-6 PMID: 8811086
  33. Importin provides a link between nuclear protein import and U snRNA export.
    Cell. 1996 Oct 4;87(1):21-32 PMID: 8858145
  34. eIF4G: a multipurpose ribosome adapter?
    Science. 1997 Jan 24;275(5299):500-1 PMID: 9019810
  35. Distribution of pre-mRNA splicing factors at sites of RNA polymerase II transcription.
    Genes Dev. 1997 May 1;11(9):1148-59 PMID: 9159396
  36. Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens.
    Nat Genet. 1997 Jul;16(3):277-82 PMID: 9207794
  37. Involvement of stress-activated protein kinase and p38/RK mitogen-activated protein kinase signaling pathways in the enhanced phosphorylation of initiation factor 4E in NIH 3T3 cells.
    J Biol Chem. 1997 Jul 11;272(28):17887-93 PMID: 9211946
  38. The role of the cap structure in RNA processing and nuclear export.
    Eur J Biochem. 1997 Jul 15;247(2):461-9 PMID: 9266685
  39. CRM1 is an export receptor for leucine-rich nuclear export signals.
    Cell. 1997 Sep 19;90(6):1051-60 PMID: 9323133
  40. eIF4G: translation's mystery factor begins to yield its secrets.
    RNA. 1997 Oct;3(10):1085-104 PMID: 9326485
  41. Participation of the nuclear cap binding complex in pre-mRNA 3' processing.
    Proc Natl Acad Sci U S A. 1997 Oct 28;94(22):11893-8 PMID: 9342333
  42. Comparison of caspase activation and subcellular localization in HL-60 and K562 cells undergoing etoposide-induced apoptosis.
    Blood. 1997 Dec 1;90(11):4283-96 PMID: 9373239
  43. 5'-Capping enzymes are targeted to pre-mRNA by binding to the phosphorylated carboxy-terminal domain of RNA polymerase II.
    Genes Dev. 1997 Dec 15;11(24):3306-18 PMID: 9407024
  44. mRNA capping enzyme is recruited to the transcription complex by phosphorylation of the RNA polymerase II carboxy-terminal domain.
    Genes Dev. 1997 Dec 15;11(24):3319-26 PMID: 9407025
  45. The guanylyltransferase domain of mammalian mRNA capping enzyme binds to the phosphorylated carboxyl-terminal domain of RNA polymerase II.
    J Biol Chem. 1998 Apr 17;273(16):9577-85 PMID: 9545288
  46. The human poly(A)-binding protein 1 shuttles between the nucleus and the cytoplasm.
    J Biol Chem. 1998 May 22;273(21):13015-21 PMID: 9582337
  47. Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1.
    Exp Cell Res. 1998 Aug 1;242(2):540-7 PMID: 9683540
  48. Leptomycin B inhibits equine infectious anemia virus Rev and feline immunodeficiency virus rev function but not the function of the hepatitis B virus posttranscriptional regulatory element.
    J Virol. 1998 Sep;72(9):7593-7 PMID: 9696859
  49. Circularization of mRNA by eukaryotic translation initiation factors.
    Mol Cell. 1998 Jul;2(1):135-40 PMID: 9702200
  50. Rotavirus RNA-binding protein NSP3 interacts with eIF4GI and evicts the poly(A) binding protein from eIF4F.
    EMBO J. 1998 Oct 1;17(19):5811-21 PMID: 9755181
  51. The association of initiation factor 4F with poly(A)-binding protein is enhanced in serum-stimulated Xenopus kidney cells.
    J Biol Chem. 1999 Jan 1;274(1):196-204 PMID: 9867830
  52. Phosphorylation of the cap-binding protein eukaryotic translation initiation factor 4E by protein kinase Mnk1 in vivo.
    Mol Cell Biol. 1999 Mar;19(3):1871-80 PMID: 10022874
  53. Identification of a nucleic acid binding domain in eukaryotic initiation factor eIFiso4G from wheat.
    J Biol Chem. 1999 Apr 9;274(15):10603-8 PMID: 10187856
  54. Involvement of CRM1, a nuclear export receptor, in mRNA export in mammalian cells and fission yeast.
    Genes Cells. 1999 May;4(5):291-7 PMID: 10421839
  55. Genetic and physical interactions involving the yeast nuclear cap-binding complex.
    Mol Cell Biol. 1999 Oct;19(10):6543-53 PMID: 10490594
  56. Luc7p, a novel yeast U1 snRNP protein with a role in 5' splice site recognition.
    Genes Dev. 1999 Sep 15;13(18):2425-38 PMID: 10500099
  57. Site-directed cross-linking of mRNA analogues to the Escherichia coli ribosome; identification of 30S ribosomal components that can be cross-linked to the mRNA at various points 5' with respect to the decoding site.
    Nucleic Acids Res. 1989 Dec 11;17(23):9889-908 PMID: 2690012
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
Published
2001-06-00
Pages
3632-41
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC86986
Subset
IM
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: [email protected]