Home LiteratureArticle Details
PMID: 9707591 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A mutated human homologue to yeast Upf1 protein has a dominant-negative effect on the decay of nonsense-containing mRNAs in mammalian cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 95 ·No. 17 ·1998-08-18 ·Pages 10009-14

Sun X, Perlick HA, Dietz HC, Maquat LE

Abstract

All eukaryotic cells analyzed have developed mechanisms to eliminate the production of mRNAs that prematurely terminate translation. The mechanisms are thought to exist to protect cells from the deleterious effects of in-frame nonsense codons that are generated by routine inefficiencies and inaccuracies in RNA metabolism such as pre-mRNA splicing. Depending on the particular mRNA and how it is produced, nonsense codons can mediate a reduction in mRNA abundance either (i) before its release from an association with nuclei into the cytoplasm, presumably but not certainly while the mRNA is being exported to the cytoplasm and translated by cytoplasmic ribosomes, or (ii) in the cytoplasm. Here, we provide evidence for a factor that functions to eliminate the production of nonsense-containing RNAs in mammalian cells. The factor, variously referred to as Rent1 (regulator of nonsense transcripts) or HUPF1 (human Upf1 protein), was identified by isolating cDNA for a human homologue to Saccharomyces cerevisiae Upf1p, which is a group I RNA helicase that functions in the nonsense-mediated decay of mRNA in yeast. Using monkey COS cells and human HeLa cells, we demonstrate that expression of human Upf1 protein harboring an arginine-to-cysteine mutation at residue 844 within the RNA helicase domain acts in a dominant-negative fashion to abrogate the decay of nonsense-containing mRNA that takes place (i) in association with nuclei or (ii) in the cytoplasm. These findings provide evidence that nonsense-mediated mRNA decay is related mechanistically in yeast and in mammalian cells, regardless of the cellular site of decay.

MeSH Terms
Animals Base Sequence COS Cells Cell Nucleus/metabolism Chlorocebus aethiops Codon, Nonsense Cytoplasm/metabolism DNA Primers/genetics DNA, Complementary/genetics Fungal Proteins/genetics Gene Expression HeLa Cells Humans Mutation RNA Helicases RNA, Messenger/genetics,metabolism Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Species Specificity Trans-Activators/genetics Transfection
Chemicals
Codon, Nonsense DNA Primers DNA, Complementary Fungal Proteins RNA, Messenger Saccharomyces cerevisiae Proteins Trans-Activators NAM7 protein, S cerevisiae RNA Helicases UPF1 protein, human
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Sun X
Roswell Park Cancer Institute, Department of Genetics, Elm and Carlton Streets, Buffalo, NY 14263, USA.
Perlick H A
Dietz H C
Maquat L E
References (44)
44 references, click to expand
  1. NAM7 nuclear gene encodes a novel member of a family of helicases with a Zn-ligand motif and is involved in mitochondrial functions in Saccharomyces cerevisiae.
    J Mol Biol. 1992 Apr 5;224(3):575-87 PMID: 1314899
  2. Gene products that promote mRNA turnover in Saccharomyces cerevisiae.
    Mol Cell Biol. 1992 May;12(5):2165-77 PMID: 1569946
  3. The product of the yeast UPF1 gene is required for rapid turnover of mRNAs containing a premature translational termination codon.
    Genes Dev. 1991 Dec;5(12A):2303-14 PMID: 1748286
  4. Development of the primary antibody repertoire.
    Science. 1987 Nov 20;238(4830):1079-87 PMID: 3317825
  5. Identification of an additional gene required for eukaryotic nonsense mRNA turnover.
    Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10354-8 PMID: 7479783
  6. When cells stop making sense: effects of nonsense codons on RNA metabolism in vertebrate cells.
    RNA. 1995 Jul;1(5):453-65 PMID: 7489507
  7. Purification and characterization of the Upf1 protein: a factor involved in translation and mRNA degradation.
    RNA. 1995 Aug;1(6):610-23 PMID: 7489520
  8. A regulatory mechanism that detects premature nonsense codons in T-cell receptor transcripts in vivo is reversed by protein synthesis inhibitors in vitro.
    J Biol Chem. 1995 Dec 1;270(48):28995-9003 PMID: 7499432
  9. The majority of yeast UPF1 co-localizes with polyribosomes in the cytoplasm.
    Mol Biol Cell. 1995 May;6(5):611-25 PMID: 7545033
  10. A genetic screen identifies cellular factors involved in retroviral -1 frameshifting.
    Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6587-91 PMID: 7604038
  11. Characterization of cis-acting sequences and decay intermediates involved in nonsense-mediated mRNA turnover.
    Mol Cell Biol. 1995 Feb;15(2):809-23 PMID: 7823948
  12. Evidence for degradation of mRNA encoding alpha-L-iduronidase in Hurler fibroblasts with premature termination alleles.
    Cell Mol Biol (Noisy-le-grand). 1994 Nov;40(7):999-1005 PMID: 7849567
  13. Identification and characterization of genes that are required for the accelerated degradation of mRNAs containing a premature translational termination codon.
    Genes Dev. 1995 Feb 15;9(4):423-36 PMID: 7883167
  14. Identification of a novel component of the nonsense-mediated mRNA decay pathway by use of an interacting protein screen.
    Genes Dev. 1995 Feb 15;9(4):437-54 PMID: 7883168
  15. Nuclear degradation of nonsense mutated beta-globin mRNA: a post-transcriptional mechanism to protect heterozygotes from severe clinical manifestations of beta-thalassemia?
    Nucleic Acids Res. 1995 Feb 11;23(3):413-8 PMID: 7885837
  16. 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
  17. Mammalian nonsense codons can be cis effectors of nuclear mRNA half-life.
    Mol Cell Biol. 1994 Dec;14(12):8219-28 PMID: 7969159
  18. Nonsense-mediated mRNA decay in yeast.
    Prog Nucleic Acid Res Mol Biol. 1994;47:271-98 PMID: 8016322
  19. Introns are cis effectors of the nonsense-codon-mediated reduction in nuclear mRNA abundance.
    Mol Cell Biol. 1994 Sep;14(9):6317-25 PMID: 8065363
  20. Nonsense but not missense mutations can decrease the abundance of nuclear mRNA for the mouse major urinary protein, while both types of mutations can facilitate exon skipping.
    Mol Cell Biol. 1994 Sep;14(9):6326-36 PMID: 8065364
  21. Stabilization and ribosome association of unspliced pre-mRNAs in a yeast upf1- mutant.
    Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7034-8 PMID: 8346213
  22. T cell receptor-beta mRNA splicing: regulation of unusual splicing intermediates.
    Mol Cell Biol. 1993 Mar;13(3):1686-96 PMID: 8441406
  23. Nonsense codons can reduce the abundance of nuclear mRNA without affecting the abundance of pre-mRNA or the half-life of cytoplasmic mRNA.
    Mol Cell Biol. 1993 Mar;13(3):1892-902 PMID: 8441420
  24. Spatial consequences of defective processing of specific yeast mRNAs revealed by fluorescent in situ hybridization.
    RNA. 1995 Dec;1(10):1071-8 PMID: 8595562
  25. Evidence that the decay of nucleus-associated nonsense mRNA for human triosephosphate isomerase involves nonsense codon recognition after splicing.
    RNA. 1996 Mar;2(3):235-43 PMID: 8608447
  26. Knowing when not to stop: selenocysteine incorporation in eukaryotes.
    Trends Biochem Sci. 1996 Jun;21(6):203-8 PMID: 8744353
  27. Defects in RNA splicing and the consequence of shortened translational reading frames.
    Am J Hum Genet. 1996 Aug;59(2):279-86 PMID: 8755945
  28. An essential component of the decapping enzyme required for normal rates of mRNA turnover.
    Nature. 1996 Aug 15;382(6592):642-6 PMID: 8757137
  29. Genetic and biochemical characterization of mutations in the ATPase and helicase regions of the Upf1 protein.
    Mol Cell Biol. 1996 Oct;16(10):5477-90 PMID: 8816461
  30. Identification and characterization of mutations in the UPF1 gene that affect nonsense suppression and the formation of the Upf protein complex but not mRNA turnover.
    Mol Cell Biol. 1996 Oct;16(10):5491-506 PMID: 8816462
  31. Mammalian orthologues of a yeast regulator of nonsense transcript stability.
    Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10928-32 PMID: 8855285
  32. Mof4-1 is an allele of the UPF1/IFS2 gene which affects both mRNA turnover and -1 ribosomal frameshifting efficiency.
    EMBO J. 1996 Oct 15;15(20):5726-36 PMID: 8896465
  33. A splicing-dependent regulatory mechanism that detects translation signals.
    EMBO J. 1996 Nov 1;15(21):5965-75 PMID: 8918474
  34. Making sense of nonsense in yeast.
    Trends Biochem Sci. 1996 Nov;21(11):433-8 PMID: 8987399
  35. Upf1p, Nmd2p, and Upf3p are interacting components of the yeast nonsense-mediated mRNA decay pathway.
    Mol Cell Biol. 1997 Mar;17(3):1580-94 PMID: 9032286
  36. Evidence that translation reinitiation abrogates nonsense-mediated mRNA decay in mammalian cells.
    EMBO J. 1997 Feb 17;16(4):826-33 PMID: 9049311
  37. Cloning and characterization of HUPF1, a human homolog of the Saccharomyces cerevisiae nonsense mRNA-reducing UPF1 protein.
    Nucleic Acids Res. 1997 Feb 15;25(4):814-21 PMID: 9064659
  38. T cell receptor (TCR) mini-gene mRNA expression regulated by nonsense codons: a nuclear-associated translation-like mechanism.
    J Exp Med. 1997 Mar 17;185(6):985-92 PMID: 9091590
  39. Relationship between yeast polyribosomes and Upf proteins required for nonsense mRNA decay.
    J Biol Chem. 1997 Aug 29;272(35):22163-72 PMID: 9268361
  40. The presence of an intron within the rat gene for selenium-dependent glutathione peroxidase 1 is not required to protect nuclear RNA from UGA-mediated decay.
    RNA. 1997 Dec;3(12):1369-73 PMID: 9404888
  41. Selenium deficiency reduces the abundance of mRNA for Se-dependent glutathione peroxidase 1 by a UGA-dependent mechanism likely to be nonsense codon-mediated decay of cytoplasmic mRNA.
    Mol Cell Biol. 1998 May;18(5):2932-9 PMID: 9566912
  42. ATP is a cofactor of the Upf1 protein that modulates its translation termination and RNA binding activities.
    RNA. 1998 Feb;4(2):205-14 PMID: 9570320
  43. 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
  44. Intron function in the nonsense-mediated decay of beta-globin mRNA: indications that pre-mRNA splicing in the nucleus can influence mRNA translation in the cytoplasm.
    RNA. 1998 Jul;4(7):801-15 PMID: 9671053
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.95.17.10009
Published
1998-08-18
Pages
10009-14
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC21452
Subset
IM
Grants
NIDDK NIH HHS · DK33933 · United States
NIGMS NIH HHS · GM52822 · United States
NIGMS NIH HHS · GM55239 · United States
External Links
PubMed source DOI: 10.1073/pnas.95.17.10009 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