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PMID: 7610048 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Autoregulation of poly(A)-binding protein synthesis in vitro.

Nucleic acids research ·Vol. 23 ·No. 12 ·1995-06-25 ·Pages 2198-205

de Melo Neto OP, Standart N, Martins de Sa C

Abstract

The poly(A)-binding protein (PABP), in a complex with the 3'poly(A) tail of eukaryotic mRNAs, plays important roles in the control of translation and message stability. All known examples of PABP mRNAs contain an extensive A-rich sequence in their 5' untranslated regions. Studies in mammalian cells undergoing growth stimulation or terminal differentiation indicate that PABP expression is regulated at the translational level. Here we examine the hypothesis that synthesis of the PABP is autogenously controlled. We show that the endogenous inactive PABP mRNA in rabbit reticulocytes can be specifically stimulated by addition of low concentrations of poly(A) and that this stimulation is also observed with in vitro transcribed human PABP mRNA. By deleting the A-rich region from the leader of human PABP mRNA and adding it upstream of the initiator AUG in a reporter mRNA we show that the adenylate tract is sufficient and necessary for mRNA repression and poly(A)-mediated activation in the reticulocyte cell-free system. UV cross-linking experiments demonstrate that the leader adenylate tract binds PABP. Furthermore, addition of recombinant GST-PABP to the cell-free system represses translation of mRNAs containing the A-rich sequence in their 5'UTR, but has no effect on control mRNA. We thus conclude that in vitro PABP binding to the A-rich sequence in the 5' UTR of PABP mRNA represses its own synthesis.

MeSH Terms
Adenosine Monophosphate/chemistry,metabolism Animals Base Sequence Cell-Free System Gene Expression Regulation/drug effects Humans Molecular Sequence Data Poly A/pharmacology Poly(A)-Binding Proteins Protein Biosynthesis/drug effects RNA, Messenger/metabolism RNA-Binding Proteins/biosynthesis,genetics Rabbits Receptors, Cytoplasmic and Nuclear Recombinant Proteins/pharmacology Reticulocytes/chemistry,metabolism Triticum
Chemicals
Poly(A)-Binding Proteins RNA, Messenger RNA-Binding Proteins Receptors, Cytoplasmic and Nuclear Recombinant Proteins Poly A Adenosine Monophosphate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
de Melo Neto O P
Department of Biochemistry, University of Cambridge, UK.
Standart N
Martins de Sa C
References (52)
52 references, click to expand
  1. Identification of an RNA binding site for human thymidylate synthase.
    Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):517-21 PMID: 8421684
  2. A single domain of yeast poly(A)-binding protein is necessary and sufficient for RNA binding and cell viability.
    Mol Cell Biol. 1987 Sep;7(9):3268-76 PMID: 3313012
  3. Drosophila melanogaster poly(A)-binding protein: cDNA cloning reveals an unusually long 3'-untranslated region of the mRNA, also present in other eukaryotic species.
    Gene. 1993 Apr 30;126(2):295-6 PMID: 8482550
  4. Specific binding of human dihydrofolate reductase protein to dihydrofolate reductase messenger RNA in vitro.
    Biochemistry. 1993 May 11;32(18):4756-60 PMID: 8490020
  5. Differential organ-specific expression of three poly(A)-binding-protein genes from Arabidopsis thaliana.
    Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6686-90 PMID: 8341686
  6. A cytoplasmic 57-kDa protein that is required for translation of picornavirus RNA by internal ribosomal entry is identical to the nuclear pyrimidine tract-binding protein.
    Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7642-6 PMID: 8395052
  7. Ribosomal protein L32 of Saccharomyces cerevisiae regulates both splicing and translation of its own transcript.
    J Biol Chem. 1993 Sep 15;268(26):19669-74 PMID: 8366109
  8. The involvement of a spliceosome component in internal initiation of human rhinovirus RNA translation.
    J Gen Virol. 1993 Sep;74 ( Pt 9):1775-88 PMID: 8397279
  9. Cytoplasmic protein binds in vitro to a highly conserved sequence in the 5' untranslated region of ferritin heavy- and light-subunit mRNAs.
    Proc Natl Acad Sci U S A. 1988 Apr;85(7):2171-5 PMID: 3127826
  10. The poly(A)-binding protein facilitates in vitro translation of poly(A)-rich mRNA.
    Eur J Biochem. 1988 Oct 1;176(3):521-6 PMID: 2901956
  11. The poly(A)-poly(A)-binding protein complex is a major determinant of mRNA stability in vitro.
    Mol Cell Biol. 1989 Feb;9(2):659-70 PMID: 2565532
  12. On the fidelity of mRNA translation in the nuclease-treated rabbit reticulocyte lysate system.
    Nucleic Acids Res. 1989 Apr 25;17(8):3129-44 PMID: 2726454
  13. Expression of the poly(A)-binding protein during development of Xenopus laevis.
    Mol Cell Biol. 1989 Jun;9(6):2756-60 PMID: 2761544
  14. The poly(A) binding protein is required for poly(A) shortening and 60S ribosomal subunit-dependent translation initiation.
    Cell. 1989 Sep 8;58(5):857-67 PMID: 2673535
  15. Poly(A), poly(A) binding protein and the regulation of mRNA stability.
    Trends Biochem Sci. 1989 Sep;14(9):373-7 PMID: 2688202
  16. mRNA poly(A) tail, a 3' enhancer of translational initiation.
    Mol Cell Biol. 1990 Jul;10(7):3441-55 PMID: 1972543
  17. Do the poly(A) tail and 3' untranslated region control mRNA translation?
    Cell. 1990 Jul 13;62(1):15-24 PMID: 1973075
  18. The A- and B-type cyclin associated cdc2 kinases in Xenopus turn on and off at different times in the cell cycle.
    EMBO J. 1990 Sep;9(9):2865-75 PMID: 2143983
  19. The Xenopus laevis poly(A) binding protein is composed of multiple functionally independent RNA binding domains.
    EMBO J. 1990 Nov;9(11):3699-705 PMID: 2209558
  20. Tales of poly(A): a review.
    Gene. 1990 Jul 16;91(2):151-8 PMID: 1976572
  21. Translational repression by a complex between the iron-responsive element of ferritin mRNA and its specific cytoplasmic binding protein is position-dependent in vivo.
    EMBO J. 1990 Dec;9(12):4127-33 PMID: 1701143
  22. A conserved element in the leader mediates post-meiotic translation as well as cytoplasmic polyadenylation of a Drosophila spermatocyte mRNA.
    EMBO J. 1990 Dec;9(13):4519-25 PMID: 2124974
  23. The multiple RNA-binding domains of the mRNA poly(A)-binding protein have different RNA-binding activities.
    Mol Cell Biol. 1991 Jul;11(7):3419-24 PMID: 1675426
  24. The role of poly(A) in the translation and stability of mRNA.
    Curr Opin Cell Biol. 1990 Dec;2(6):1092-8 PMID: 2099802
  25. Characterization of cDNAs encoding the polypyrimidine tract-binding protein.
    Genes Dev. 1991 Jul;5(7):1224-36 PMID: 1906035
  26. Characterization and molecular cloning of polypyrimidine tract-binding protein: a component of a complex necessary for pre-mRNA splicing.
    Genes Dev. 1991 Jul;5(7):1237-51 PMID: 1906036
  27. Sequences mediating the translation of mouse S16 ribosomal protein mRNA during myoblast differentiation and in vitro and possible control points for the in vitro translation.
    Genes Dev. 1991 Sep;5(9):1723-36 PMID: 1885008
  28. Autoregulation of human thymidylate synthase messenger RNA translation by thymidylate synthase.
    Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8977-81 PMID: 1924359
  29. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency.
    Genes Dev. 1991 Nov;5(11):2108-16 PMID: 1682219
  30. Identification of proteins associating with poly(A)-binding-protein mRNA.
    Eur J Biochem. 1992 Mar 1;204(2):733-43 PMID: 1541286
  31. Position is the critical determinant for function of iron-responsive elements as translational regulators.
    Mol Cell Biol. 1992 May;12(5):1959-66 PMID: 1569933
  32. Visualizing mRNA expression in plant protoplasts: factors influencing efficient mRNA uptake and translation.
    Plant Cell. 1989 Mar;1(3):301-11 PMID: 2535505
  33. Translation initiation requires the PAB-dependent poly(A) ribonuclease in yeast.
    Cell. 1992 Sep 18;70(6):961-73 PMID: 1339314
  34. Bacteriophage and spliceosomal proteins function as position-dependent cis/trans repressors of mRNA translation in vitro.
    Nucleic Acids Res. 1992 Nov 11;20(21):5555-64 PMID: 1454520
  35. Regulation of translation in eukaryotic systems.
    Annu Rev Cell Biol. 1992;8:197-225 PMID: 1335743
  36. Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors.
    Nucleic Acids Res. 1993 Dec 25;21(25):5803-16 PMID: 8290338
  37. The human U1A snRNP protein regulates polyadenylation via a direct interaction with poly(A) polymerase.
    Cell. 1994 Feb 11;76(3):531-41 PMID: 8313473
  38. Gene regulation in Drosophila spermatogenesis: analysis of protein binding at the translational control element TCE.
    Dev Genet. 1993;14(6):449-59 PMID: 8111973
  39. Translation of 15-lipoxygenase mRNA is inhibited by a protein that binds to a repeated sequence in the 3' untranslated region.
    EMBO J. 1994 Mar 15;13(6):1476-81 PMID: 8137829
  40. Vertebrate mRNAs with a 5'-terminal pyrimidine tract are candidates for translational repression in quiescent cells: characterization of the translational cis-regulatory element.
    Mol Cell Biol. 1994 Jun;14(6):3822-33 PMID: 8196625
  41. Poly(A) binds to initiation factors and increases cap-dependent translation in vitro.
    J Biol Chem. 1994 Jun 24;269(25):17166-73 PMID: 8006024
  42. Conserved structures and diversity of functions of RNA-binding proteins.
    Science. 1994 Jul 29;265(5172):615-21 PMID: 8036511
  43. Proteins binding to 5' untranslated region sites: a general mechanism for translational regulation of mRNAs in human and yeast cells.
    Mol Cell Biol. 1994 Sep;14(9):5898-909 PMID: 8065323
  44. Iron regulatory protein prevents binding of the 43S translation pre-initiation complex to ferritin and eALAS mRNAs.
    EMBO J. 1994 Aug 15;13(16):3882-91 PMID: 8070415
  45. Characterization of a Trypanosoma cruzi poly(A)-binding protein and its genes.
    Mol Biochem Parasitol. 1994 Oct;67(2):301-12 PMID: 7870134
  46. Messenger RNA for the 73,000-dalton poly(A)-binding protein occurs as translationally repressed mRNP in duck reticulocytes.
    J Biol Chem. 1983 Feb 10;258(3):1387-90 PMID: 6822518
  47. The translation of the messenger for the poly(A)-binding protein-associated with translated mRNA is suppressed. A case of cytoplasmic repression in duck erythroblasts.
    FEBS Lett. 1983 Oct 3;162(1):25-32 PMID: 6617892
  48. Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA.
    Methods Enzymol. 1983;96:50-74 PMID: 6656641
  49. A single gene from yeast for both nuclear and cytoplasmic polyadenylate-binding proteins: domain structure and expression.
    Cell. 1986 Jun 20;45(6):827-35 PMID: 3518950
  50. Translational control of mRNA expression during the early mitogenic response in Swiss mouse 3T3 cells: identification of specific proteins.
    J Cell Biol. 1986 Dec;103(6 Pt 1):2137-44 PMID: 3782293
  51. Human mRNA polyadenylate binding protein: evolutionary conservation of a nucleic acid binding motif.
    Nucleic Acids Res. 1987 Jun 25;15(12):4771-87 PMID: 2885805
  52. Translational regulation by mRNA/protein interactions in eukaryotic cells: ferritin and beyond.
    Bioessays. 1993 Feb;15(2):85-90 PMID: 8471060
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/23.12.2198
Published
1995-06-25
Pages
2198-205
Language
English
Region
England
NLM ID
0411011
PMCID
PMC307008
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/23.12.2198 PMC Full Text View journal details
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