Home LiteratureArticle Details
PMID: 9784497 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

mRNA stabilization by poly(A) binding protein is independent of poly(A) and requires translation.

Genes & development ·Vol. 12 ·No. 20 ·1998-10-15 ·Pages 3226-35

Coller JM, Gray NK, Wickens MP

Abstract

Translation and mRNA stability are enhanced by the presence of a poly(A) tail. In vivo, the tail interacts with a conserved polypeptide, poly(A) binding protein (Pab1p). To examine Pab1p function in vivo, we have tethered Pab1p to the 3' UTR of reporter mRNAs by fusing it to MS2 coat protein and placing MS2 binding sites in the 3' UTR of the reporter. This strategy allows us to uncouple Pab1p function from its RNA binding activity. We show that mRNAs that lack a poly(A) tail in vivo are stabilized by Pab1p, and that the portions of Pab1p required for stabilization are genetically distinct from those required for poly(A) binding. In addition, stabilization by Pab1p requires ongoing translation of the mRNA. We conclude that the primary, or sole, function of poly(A) with respect to mRNA stability is simply to bring Pab1p to the mRNA, and that mRNA stabilization is an intrinsic property of Pab1p. The approach we describe may be useful in identifying and assaying 3' UTR regulatory proteins, as it uncouples analysis of function from RNA binding.

MeSH Terms
3' Untranslated Regions/metabolism Binding Sites/genetics Capsid/genetics,metabolism Capsid Proteins Peptide Chain Elongation, Translational/genetics Poly A/metabolism Poly(A)-Binding Proteins Protein Binding/genetics Protein Biosynthesis/genetics,physiology RNA, Messenger/genetics,metabolism RNA-Binding Proteins/genetics,metabolism,physiology Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae
Chemicals
3' Untranslated Regions Capsid Proteins Poly(A)-Binding Proteins RNA, Messenger RNA-Binding Proteins Recombinant Fusion Proteins Poly A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Coller J M
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706-1544 USA.
Gray N K
Wickens M P
References (48)
48 references, click to expand
  1. Deadenylation of the unstable mRNA encoded by the yeast MFA2 gene leads to decapping followed by 5'-->3' digestion of the transcript.
    Genes Dev. 1994 Apr 1;8(7):855-66 PMID: 7926773
  2. Defects in mRNA 3'-end formation, transcription initiation, and mRNA transport associated with the yeast mutation prp20: possible coupling of mRNA processing and chromatin structure.
    Genes Dev. 1992 Oct;6(10):1914-26 PMID: 1398069
  3. Formation of brome mosaic virus RNA-dependent RNA polymerase in yeast requires coexpression of viral proteins and viral RNA.
    Proc Natl Acad Sci U S A. 1995 May 23;92(11):4892-6 PMID: 7761419
  4. mRNA decay mediated by two distinct AU-rich elements from c-fos and granulocyte-macrophage colony-stimulating factor transcripts: different deadenylation kinetics and uncoupling from translation.
    Mol Cell Biol. 1995 Oct;15(10):5777-88 PMID: 7565731
  5. Multiple functions for the poly(A)-binding protein in mRNA decapping and deadenylation in yeast.
    Genes Dev. 1995 Oct 1;9(19):2421-32 PMID: 7557393
  6. AU-rich elements: characterization and importance in mRNA degradation.
    Trends Biochem Sci. 1995 Nov;20(11):465-70 PMID: 8578590
  7. Selection of RNA-binding peptides in vivo.
    Nature. 1996 Mar 14;380(6570):175-9 PMID: 8600395
  8. Xenopus poly(A) binding protein: functional domains in RNA binding and protein-protein interaction.
    J Mol Biol. 1996 Feb 16;256(1):20-30 PMID: 8609610
  9. Role of polyadenylation in nucleocytoplasmic transport of mRNA.
    Mol Cell Biol. 1996 Apr;16(4):1534-42 PMID: 8657127
  10. A three-hybrid system to detect RNA-protein interactions in vivo.
    Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8496-501 PMID: 8710898
  11. An essential component of the decapping enzyme required for normal rates of mRNA turnover.
    Nature. 1996 Aug 15;382(6592):642-6 PMID: 8757137
  12. Interrelationships of the pathways of mRNA decay and translation in eukaryotic cells.
    Annu Rev Biochem. 1996;65:693-739 PMID: 8811193
  13. Mutations in trans-acting factors affecting mRNA decapping in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Oct;16(10):5830-8 PMID: 8816497
  14. Mechanisms and control of mRNA turnover in Saccharomyces cerevisiae.
    Microbiol Rev. 1996 Mar;60(1):233-49 PMID: 8852902
  15. Association of the yeast poly(A) tail binding protein with translation initiation factor eIF-4G.
    EMBO J. 1996 Dec 16;15(24):7168-77 PMID: 9003792
  16. 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
  17. Eucaryotic RNA polymerase conditional mutant that rapidly ceases mRNA synthesis.
    Mol Cell Biol. 1987 May;7(5):1602-11 PMID: 3299050
  18. 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
  19. An RNA mutation that increases the affinity of an RNA-protein interaction.
    Nucleic Acids Res. 1987 Dec 23;15(24):10483-93 PMID: 3697094
  20. The SPA2 protein of yeast localizes to sites of cell growth.
    J Cell Biol. 1989 Apr;108(4):1419-29 PMID: 2647769
  21. 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
  22. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites.
    Gene. 1988 Dec 30;74(2):527-34 PMID: 3073106
  23. 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
  24. Identification and comparison of stable and unstable mRNAs in Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 May;10(5):2269-84 PMID: 2183028
  25. 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
  26. Purification of RNA and RNA-protein complexes by an R17 coat protein affinity method.
    Nucleic Acids Res. 1990 Nov 25;18(22):6587-94 PMID: 1701242
  27. Specific interaction between RNA phage coat proteins and RNA.
    Prog Nucleic Acid Res Mol Biol. 1991;40:185-220 PMID: 2031083
  28. Life and death in the cytoplasm: messages from the 3' end.
    Curr Opin Genet Dev. 1997 Apr;7(2):220-32 PMID: 9115434
  29. 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
  30. RNA recognition motif 2 of yeast Pab1p is required for its functional interaction with eukaryotic translation initiation factor 4G.
    Mol Cell Biol. 1998 Jan;18(1):51-7 PMID: 9418852
  31. 3'-UTR-dependent deadenylation by the yeast poly(A) nuclease.
    Genes Dev. 1992 Nov;6(11):2088-99 PMID: 1358757
  32. Mutations affecting stability and deadenylation of the yeast MFA2 transcript.
    Genes Dev. 1992 Nov;6(11):2100-11 PMID: 1427074
  33. 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
  34. The protein Sex-lethal antagonizes the splicing factor U2AF to regulate alternative splicing of transformer pre-mRNA.
    Nature. 1993 Mar 11;362(6416):171-5 PMID: 7680770
  35. A turnover pathway for both stable and unstable mRNAs in yeast: evidence for a requirement for deadenylation.
    Genes Dev. 1993 Aug;7(8):1632-43 PMID: 8393418
  36. mRNA surveillance by the Caenorhabditis elegans smg genes.
    Genes Dev. 1993 Oct;7(10):1885-97 PMID: 8104846
  37. Recombinant iron-regulatory factor functions as an iron-responsive-element-binding protein, a translational repressor and an aconitase. A functional assay for translational repression and direct demonstration of the iron switch.
    Eur J Biochem. 1993 Dec 1;218(2):657-67 PMID: 8269957
  38. The half-life of c-myc mRNA in growing and serum-stimulated cells: influence of the coding and 3' untranslated regions and role of ribosome translocation.
    Mol Cell Biol. 1994 Mar;14(3):2119-28 PMID: 8114742
  39. Differential effects of translational inhibition in cis and in trans on the decay of the unstable yeast MFA2 mRNA.
    J Biol Chem. 1994 Apr 1;269(13):9687-92 PMID: 8144558
  40. 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
  41. Premature translational termination triggers mRNA decapping.
    Nature. 1994 Aug 18;370(6490):578-81 PMID: 8052314
  42. A comparison of mammalian and yeast pre-mRNA 3'-end processing.
    Curr Opin Cell Biol. 1997 Jun;9(3):329-36 PMID: 9159082
  43. Differential effects of aromatic and charged residue substitutions in the RNA binding domains of the yeast poly(A)-binding protein.
    J Mol Biol. 1997 May 30;269(1):67-81 PMID: 9193001
  44. Yeast Pab1 interacts with Rna15 and participates in the control of the poly(A) tail length in vitro.
    Mol Cell Biol. 1997 Jul;17(7):3694-701 PMID: 9199303
  45. Starting at the beginning, middle, and end: translation initiation in eukaryotes.
    Cell. 1997 Jun 13;89(6):831-8 PMID: 9200601
  46. The major yeast poly(A)-binding protein is associated with cleavage factor IA and functions in premessenger RNA 3'-end formation.
    Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):7897-902 PMID: 9223284
  47. Kinetics of hairpin ribozyme cleavage in yeast.
    RNA. 1997 Sep;3(9):961-73 PMID: 9292496
  48. Turnover mechanisms of the stable yeast PGK1 mRNA.
    Mol Cell Biol. 1995 Apr;15(4):2145-56 PMID: 7891709
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.12.20.3226
Published
1998-10-15
Pages
3226-35
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC317214
Subset
IM
Grants
NIGMS NIH HHS · GM50942 · United States
External Links
PubMed source DOI: 10.1101/gad.12.20.3226 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