Home LiteratureArticle Details
PMID: 10373504 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

An mRNA stability complex functions with poly(A)-binding protein to stabilize mRNA in vitro.

Molecular and cellular biology ·Vol. 19 ·No. 7 ·1999-07-00 ·Pages 4552-60

Wang Z, Day N, Trifillis P, Kiledjian M

Abstract

The stable globin mRNAs provide an ideal system for studying the mechanism governing mammalian mRNA turnover. alpha-Globin mRNA stability is dictated by sequences in the 3' untranslated region (3'UTR) which form a specific ribonucleoprotein complex (alpha-complex) whose presence correlates with mRNA stability. One of the major protein components within this complex is a family of two polycytidylate-binding proteins, alphaCP1 and alphaCP2. Using an in vitro-transcribed and polyadenylated alpha-globin 3'UTR, we have devised an in vitro mRNA decay assay which reproduces the alpha-complex-dependent mRNA stability observed in cells. Incubation of the RNA with erythroleukemia K562 cytosolic extract results in deadenylation with distinct intermediates containing a periodicity of approximately 30 nucleotides, which is consistent with the binding of poly(A)-binding protein (PABP) monomers. Disruption of the alpha-complex by sequestration of alphaCP1 and alphaCP2 enhances deadenylation and decay of the mRNA, while reconstitution of the alpha-complex stabilizes the mRNA. Similarly, PABP is also essential for the stability of mRNA in vitro, since rapid deadenylation resulted upon its depletion. An RNA-dependent interaction between alphaCP1 and alphaCP2 with PABP suggests that the alpha-complex can directly interact with PABP. Therefore, the alpha-complex is an mRNA stability complex in vitro which could function at least in part by interacting with PABP.

MeSH Terms
3' Untranslated Regions Animals Cattle DNA-Binding Proteins Globins/genetics HeLa Cells Heterogeneous-Nuclear Ribonucleoproteins Humans K562 Cells Poly(A)-Binding Proteins RNA, Messenger/metabolism RNA-Binding Proteins/genetics,metabolism Transcription Factors
Chemicals
3' Untranslated Regions DNA-Binding Proteins Heterogeneous-Nuclear Ribonucleoproteins PCBP1 protein, human Poly(A)-Binding Proteins RNA, Messenger RNA-Binding Proteins Transcription Factors Globins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wang Z
Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, New Jersey 08854-8082, USA.
Day N
Trifillis P
Kiledjian M
References (61)
61 references, click to expand
  1. 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
  2. Cytoplasmic regulation of mRNA function: the importance of the 3' untranslated region.
    Cell. 1993 Jul 16;74(1):9-14 PMID: 7687524
  3. Four highly stable eukaryotic mRNAs assemble 3' untranslated region RNA-protein complexes sharing cis and trans components.
    Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2410-4 PMID: 9122208
  4. Poly(A) tail shortening by a mammalian poly(A)-specific 3'-exoribonuclease.
    J Biol Chem. 1997 Apr 18;272(16):10448-56 PMID: 9099687
  5. Messenger RNA deadenylylation precedes decapping in mammalian cells.
    Proc Natl Acad Sci U S A. 1997 May 27;94(11):5628-33 PMID: 9159123
  6. mRNA silencing in erythroid differentiation: hnRNP K and hnRNP E1 regulate 15-lipoxygenase translation from the 3' end.
    Cell. 1997 May 16;89(4):597-606 PMID: 9160751
  7. The role of mRNA stability in the control of globin gene expression.
    Prog Nucleic Acid Res Mol Biol. 1997;57:249-87 PMID: 9175436
  8. Requirement of poly(rC) binding protein 2 for translation of poliovirus RNA.
    J Virol. 1997 Aug;71(8):6243-6 PMID: 9223526
  9. Identification of AUF1 (heterogeneous nuclear ribonucleoprotein D) as a component of the alpha-globin mRNA stability complex.
    Mol Cell Biol. 1997 Aug;17(8):4870-6 PMID: 9234743
  10. Two functional complexes formed by KH domain containing proteins with the 5' noncoding region of poliovirus RNA.
    RNA. 1997 Aug;3(8):882-92 PMID: 9257647
  11. Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE-containing mRNAs.
    EMBO J. 1998 Jun 15;17(12):3448-60 PMID: 9628880
  12. RNA stabilization by the AU-rich element binding protein, HuR, an ELAV protein.
    EMBO J. 1998 Jun 15;17(12):3461-70 PMID: 9628881
  13. A rule for termination-codon position within intron-containing genes: when nonsense affects RNA abundance.
    Trends Biochem Sci. 1998 Jun;23(6):198-9 PMID: 9644970
  14. The deadenylating nuclease (DAN) is involved in poly(A) tail removal during the meiotic maturation of Xenopus oocytes.
    EMBO J. 1998 Sep 15;17(18):5427-37 PMID: 9736620
  15. mRNA stabilization by poly(A) binding protein is independent of poly(A) and requires translation.
    Genes Dev. 1998 Oct 15;12(20):3226-35 PMID: 9784497
  16. Pbp1p, a factor interacting with Saccharomyces cerevisiae poly(A)-binding protein, regulates polyadenylation.
    Mol Cell Biol. 1998 Dec;18(12):7383-96 PMID: 9819425
  17. Characterization of c-myc 3' to 5' mRNA decay activities in an in vitro system.
    J Biol Chem. 1998 Dec 25;273(52):34770-4 PMID: 9857001
  18. ELAV proteins stabilize deadenylated intermediates in a novel in vitro mRNA deadenylation/degradation system.
    Genes Dev. 1999 Jan 15;13(2):188-201 PMID: 9925643
  19. Purification and RNA binding properties of the polycytidylate-binding proteins alphaCP1 and alphaCP2.
    Methods. 1999 Jan;17(1):84-91 PMID: 10075886
  20. 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
  21. Size heterogeneity of polyadenylate sequences in mouse globin messenger RNA.
    J Mol Biol. 1974 Jun 25;86(2):363-71 PMID: 4415788
  22. Poly(A) size class distribution in globin mRNAs as a function of time.
    Nature. 1975 Feb 27;253(5494):749-51 PMID: 1113872
  23. Different lifetimes of reticulocyte messenger RNA.
    Cell. 1976 Jan;7(1):59-65 PMID: 949743
  24. Stability of globin mRNA in terminally differentiating murine erythroleukemia cells.
    Cell. 1981 Feb;23(2):509-14 PMID: 6937265
  25. mRNA cap binding proteins: essential factors for initiating translation.
    Cell. 1985 Feb;40(2):223-4 PMID: 3967293
  26. Half-lives of beta and gamma globin messenger RNAs and of protein synthetic capacity in cultured human reticulocytes.
    Blood. 1985 Nov;66(5):1149-54 PMID: 4052630
  27. 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
  28. 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
  29. The c-fos transcript is targeted for rapid decay by two distinct mRNA degradation pathways.
    Genes Dev. 1989 Jan;3(1):60-72 PMID: 2496006
  30. 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
  31. A novel genetic system to detect protein-protein interactions.
    Nature. 1989 Jul 20;340(6230):245-6 PMID: 2547163
  32. Translation and a 42-nucleotide segment within the coding region of the mRNA encoded by the MAT alpha 1 gene are involved in promoting rapid mRNA decay in yeast.
    Proc Natl Acad Sci U S A. 1990 Apr;87(7):2780-4 PMID: 2181450
  33. Alpha thalassemia.
    Hemoglobin. 1989;13(7-8):685-731 PMID: 2699473
  34. Catalytic activity of vaccinia mRNA capping enzyme subunits coexpressed in Escherichia coli.
    J Biol Chem. 1990 Jul 15;265(20):11960-6 PMID: 2164022
  35. Messenger RNA turnover in cell-free extracts.
    Methods Enzymol. 1990;181:202-9 PMID: 1974321
  36. 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
  37. Two distinct destabilizing elements in the c-fos message trigger deadenylation as a first step in rapid mRNA decay.
    Genes Dev. 1991 Feb;5(2):221-31 PMID: 1899842
  38. An A + U-rich element RNA-binding factor regulates c-myc mRNA stability in vitro.
    Mol Cell Biol. 1991 May;11(5):2460-6 PMID: 1901943
  39. Messenger RNA degradation in eukaryotes.
    Cell. 1993 Aug 13;74(3):413-21 PMID: 7688664
  40. Purification, characterization, and cDNA cloning of an AU-rich element RNA-binding protein, AUF1.
    Mol Cell Biol. 1993 Dec;13(12):7652-65 PMID: 8246982
  41. 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
  42. Translational maintenance of frame: mutants of Saccharomyces cerevisiae with altered -1 ribosomal frameshifting efficiencies.
    Genetics. 1994 Jan;136(1):75-86 PMID: 8138178
  43. The mRNA poly(A)-binding protein: localization, abundance, and RNA-binding specificity.
    Exp Cell Res. 1994 Apr;211(2):400-7 PMID: 7908267
  44. Premature translational termination triggers mRNA decapping.
    Nature. 1994 Aug 18;370(6490):578-81 PMID: 8052314
  45. 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
  46. Erythroid cell-specific determinants of alpha-globin mRNA stability.
    Mol Cell Biol. 1994 Dec;14(12):8123-32 PMID: 7969150
  47. Detection and characterization of a 3' untranslated region ribonucleoprotein complex associated with human alpha-globin mRNA stability.
    Mol Cell Biol. 1995 Mar;15(3):1769-77 PMID: 7862166
  48. Decay of globin mRNA during megakaryocytic differentiation of erythroleukemic cell line is accomplished through shortening of the poly(A) tail and degradation from the 3' end of the transcript.
    Biochim Biophys Acta. 1995 Apr 4;1261(2):265-71 PMID: 7711070
  49. Erythroid cell-specific mRNA stability elements in the alpha 2-globin 3' nontranslated region.
    Mol Cell Biol. 1995 May;15(5):2457-65 PMID: 7739530
  50. Characterisation of two major cellular poly(rC)-binding human proteins, each containing three K-homologous (KH) domains.
    Eur J Biochem. 1995 Jun 1;230(2):447-53 PMID: 7607214
  51. Diversity of cytoplasmic functions for the 3' untranslated region of eukaryotic transcripts.
    Curr Opin Cell Biol. 1995 Jun;7(3):386-92 PMID: 7662369
  52. Identification of two KH domain proteins in the alpha-globin mRNP stability complex.
    EMBO J. 1995 Sep 1;14(17):4357-64 PMID: 7556077
  53. mRNA stability in mammalian cells.
    Microbiol Rev. 1995 Sep;59(3):423-50 PMID: 7565413
  54. AU-rich elements: characterization and importance in mRNA degradation.
    Trends Biochem Sci. 1995 Nov;20(11):465-70 PMID: 8578590
  55. 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
  56. Interrelationships of the pathways of mRNA decay and translation in eukaryotic cells.
    Annu Rev Biochem. 1996;65:693-739 PMID: 8811193
  57. Mechanisms and control of mRNA turnover in Saccharomyces cerevisiae.
    Microbiol Rev. 1996 Mar;60(1):233-49 PMID: 8852902
  58. The poly(A) tail inhibits the assembly of a 3'-to-5' exonuclease in an in vitro RNA stability system.
    Mol Cell Biol. 1997 Jan;17(1):398-406 PMID: 8972220
  59. Control of c-myc mRNA half-life in vitro by a protein capable of binding to a coding region stability determinant.
    Genes Dev. 1992 Apr;6(4):642-54 PMID: 1559612
  60. Primary structure and binding activity of the hnRNP U protein: binding RNA through RGG box.
    EMBO J. 1992 Jul;11(7):2655-64 PMID: 1628625
  61. Destabilization of human alpha-globin mRNA by translation anti-termination is controlled during erythroid differentiation and is paralleled by phased shortening of the poly(A) tail.
    J Biol Chem. 1997 Mar 7;272(10):6607-13 PMID: 9045690
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.7.4552
Published
1999-07-00
Pages
4552-60
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC84253
Subset
IM
Grants
NIDDK NIH HHS · R01 DK051611 · United States
NIDDK NIH HHS · DK51611 · United States
External Links
PubMed source DOI: 10.1128/MCB.19.7.4552 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: [email protected]

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