Home LiteratureArticle Details
PMID: 10445880 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Overexpression of truncated Nmd3p inhibits protein synthesis in yeast.

RNA (New York, N.Y.) ·Vol. 5 ·No. 8 ·1999-08-00 ·Pages 1055-70

Belk JP, He F, Jacobson A

Abstract

The yeast NMD3 gene was identified in a two-hybrid screen using the nonsense-mediated mRNA decay factor, Upf1p, as bait. NMD3 was shown to encode an essential, highly conserved protein that associated principally with free 60S ribosomal subunits. Overexpression of a truncated form of Nmd3p, lacking 100 C-terminal amino acids and most of its Upf1p-interacting domain, had dominant-negative effects on both cell growth and protein synthesis and promoted the formation of polyribosome half-mers. These effects were eliminated by truncation of an additional 100 amino acids from Nmd3p. Overexpression of the nmd3delta100 allele also led to increased synthesis and destabilization of some ribosomal protein mRNAs, and increased synthesis and altered processing of 35S pre-rRNA. Our data suggest that Nmd3p has a role in the formation, function, or maintenance of the 60S ribosomal subunit and may provide a link for Upf1p to 80S monosomes.

MeSH Terms
Amino Acid Sequence Blotting, Northern Cell Division/genetics Cloning, Molecular Fungal Proteins/genetics,metabolism,physiology Galactose/pharmacology Gene Expression Regulation, Fungal Genes, Dominant Models, Genetic Molecular Sequence Data Polyribosomes/metabolism Protein Biosynthesis Protein Synthesis Inhibitors/metabolism RNA Helicases/metabolism RNA, Messenger/metabolism RNA, Ribosomal/genetics,metabolism RNA-Binding Proteins Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Time Factors
Chemicals
Fungal Proteins NMD3 protein, S cerevisiae Protein Synthesis Inhibitors RNA, Messenger RNA, Ribosomal RNA-Binding Proteins Saccharomyces cerevisiae Proteins NAM7 protein, S cerevisiae RNA Helicases Galactose
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Belk J P
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester 01655-0122, USA.
He F
Jacobson A
References (56)
56 references, click to expand
  1. Turnover mechanisms of the stable yeast PGK1 mRNA.
    Mol Cell Biol. 1995 Apr;15(4):2145-56 PMID: 7891709
  2. Yeast virus propagation depends critically on free 60S ribosomal subunit concentration.
    Mol Cell Biol. 1995 May;15(5):2772-81 PMID: 7739558
  3. The majority of yeast UPF1 co-localizes with polyribosomes in the cytoplasm.
    Mol Biol Cell. 1995 May;6(5):611-25 PMID: 7545033
  4. 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
  5. Purification and characterization of the Upf1 protein: a factor involved in translation and mRNA degradation.
    RNA. 1995 Aug;1(6):610-23 PMID: 7489520
  6. Interaction between Nmd2p and Upf1p is required for activity but not for dominant-negative inhibition of the nonsense-mediated mRNA decay pathway in yeast.
    RNA. 1996 Feb;2(2):153-70 PMID: 8601282
  7. An essential component of the decapping enzyme required for normal rates of mRNA turnover.
    Nature. 1996 Aug 15;382(6592):642-6 PMID: 8757137
  8. Interrelationships of the pathways of mRNA decay and translation in eukaryotic cells.
    Annu Rev Biochem. 1996;65:693-739 PMID: 8811193
  9. 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
  10. RRP5 is required for formation of both 18S and 5.8S rRNA in yeast.
    EMBO J. 1996 Oct 15;15(20):5701-14 PMID: 8896463
  11. 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
  12. QSR1, an essential yeast gene with a genetic relationship to a subunit of the mitochondrial cytochrome bc1 complex, codes for a 60 S ribosomal subunit protein.
    J Biol Chem. 1997 May 16;272(20):13372-9 PMID: 9148960
  13. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.
    Nucleic Acids Res. 1997 Sep 1;25(17):3389-402 PMID: 9254694
  14. Relationship between yeast polyribosomes and Upf proteins required for nonsense mRNA decay.
    J Biol Chem. 1997 Aug 29;272(35):22163-72 PMID: 9268361
  15. Qsr1p, a 60S ribosomal subunit protein, is required for joining of 40S and 60S subunits.
    Mol Cell Biol. 1997 Sep;17(9):5136-45 PMID: 9271391
  16. A new nomenclature for the cytoplasmic ribosomal proteins of Saccharomyces cerevisiae.
    Nucleic Acids Res. 1997 Dec 15;25(24):4872-5 PMID: 9396790
  17. The C-terminal silencing domain of Rap1p is essential for the repression of ribosomal protein genes in response to a defect in the secretory pathway.
    Nucleic Acids Res. 1998 Feb 15;26(4):1063-9 PMID: 9461469
  18. Isolation and characterization of Dcp1p, the yeast mRNA decapping enzyme.
    EMBO J. 1998 Mar 2;17(5):1487-96 PMID: 9482745
  19. Heterologous complementation reveals that mutant alleles of QSR1 render 60S ribosomal subunits unstable and translationally inactive.
    Nucleic Acids Res. 1998 May 15;26(10):2442-8 PMID: 9580698
  20. A single amino acid substitution in yeast eIF-5A results in mRNA stabilization.
    EMBO J. 1998 May 15;17(10):2914-25 PMID: 9582285
  21. The surveillance complex interacts with the translation release factors to enhance termination and degrade aberrant mRNAs.
    Genes Dev. 1998 Jun 1;12(11):1665-77 PMID: 9620853
  22. A mutated human homologue to yeast Upf1 protein has a dominant-negative effect on the decay of nonsense-containing mRNAs in mammalian cells.
    Proc Natl Acad Sci U S A. 1998 Aug 18;95(17):10009-14 PMID: 9707591
  23. The Yeast Proteome Database (YPD): a model for the organization and presentation of genome-wide functional data.
    Nucleic Acids Res. 1999 Jan 1;27(1):69-73 PMID: 9847145
  24. NMD3 encodes an essential cytoplasmic protein required for stable 60S ribosomal subunits in Saccharomyces cerevisiae.
    Mol Cell Biol. 1999 Mar;19(3):2389-99 PMID: 10022925
  25. Linking mRNA turnover and translation: assessing the polyribosomal association of mRNA decay factors and degradative intermediates.
    Methods. 1999 Jan;17(1):28-37 PMID: 10075880
  26. 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
  27. Effect of anisomycin on the cellular level of native ribosomal subunits.
    Biochemistry. 1977 May 31;16(11):2343-8 PMID: 558799
  28. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  29. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  30. Characterization of yeast strains with conditionally expressed variants of ribosomal protein genes tcm1 and cyh2.
    Mol Cell Biol. 1985 Jan;5(1):99-108 PMID: 3885011
  31. Effects of progressive depletion of TCM1 or CYH2 mRNA on Saccharomyces cerevisiae ribosomal protein accumulation.
    Mol Cell Biol. 1986 May;6(5):1535-44 PMID: 3537704
  32. Regulation of mRNA entry into polysomes. Parameters affecting polysome size and the fraction of mRNA in polysomes.
    J Biol Chem. 1987 Aug 25;262(24):11501-6 PMID: 3624224
  33. Characterization of a 40S ribosomal subunit complex in polyribosomes of Saccharomyces cerevisiae treated with cycloheximide.
    Mol Cell Biol. 1981 Jan;1(1):51-7 PMID: 6765595
  34. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector.
    Gene. 1987;60(2-3):237-43 PMID: 3327750
  35. Depletion of Saccharomyces cerevisiae ribosomal protein L16 causes a decrease in 60S ribosomal subunits and formation of half-mer polyribosomes.
    Genes Dev. 1988 Feb;2(2):160-72 PMID: 3282992
  36. mRNA-decapping enzyme from Saccharomyces cerevisiae: purification and unique specificity for long RNA chains.
    Mol Cell Biol. 1988 May;8(5):2005-10 PMID: 2838740
  37. Elevated recombination rates in transcriptionally active DNA.
    Cell. 1989 Feb 24;56(4):619-30 PMID: 2645056
  38. A novel genetic system to detect protein-protein interactions.
    Nature. 1989 Jul 20;340(6230):245-6 PMID: 2547163
  39. The polymerase chain reaction.
    Trends Genet. 1989 Jun;5(6):185-9 PMID: 2672459
  40. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier.
    Curr Genet. 1989 Dec;16(5-6):339-46 PMID: 2692852
  41. Primer extension analysis of RNA.
    Methods Enzymol. 1989;180:347-69 PMID: 2482424
  42. Identification and comparison of stable and unstable mRNAs in Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 May;10(5):2269-84 PMID: 2183028
  43. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast.
    Methods Enzymol. 1991;194:281-301 PMID: 2005793
  44. Association of RAP1 binding sites with stringent control of ribosomal protein gene transcription in Saccharomyces cerevisiae.
    Mol Cell Biol. 1991 May;11(5):2723-35 PMID: 2017175
  45. Assembly of 60S ribosomal subunits is perturbed in temperature-sensitive yeast mutants defective in ribosomal protein L16.
    Mol Cell Biol. 1991 Nov;11(11):5681-92 PMID: 1922070
  46. The yeast IMP1 gene is allelic to GAL2.
    Mol Gen Genet. 1991 Nov;230(1-2):129-35 PMID: 1745225
  47. 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
  48. Gene products that promote mRNA turnover in Saccharomyces cerevisiae.
    Mol Cell Biol. 1992 May;12(5):2165-77 PMID: 1569946
  49. Yeast cells lacking 5'-->3' exoribonuclease 1 contain mRNA species that are poly(A) deficient and partially lack the 5' cap structure.
    Mol Cell Biol. 1993 Aug;13(8):4826-35 PMID: 8336719
  50. 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
  51. 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
  52. mRNA destabilization triggered by premature translational termination depends on at least three cis-acting sequence elements and one trans-acting factor.
    Genes Dev. 1993 Sep;7(9):1737-54 PMID: 8370523
  53. Premature translational termination triggers mRNA decapping.
    Nature. 1994 Aug 18;370(6490):578-81 PMID: 8052314
  54. 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
  55. Mechanisms of mRNA degradation in eukaryotes.
    Trends Biochem Sci. 1994 Aug;19(8):336-40 PMID: 7940679
  56. 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
Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1017/s1355838299990027
Published
1999-08-00
Pages
1055-70
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1369829
Subset
IM
Grants
NIGMS NIH HHS · R01 GM027757 · United States
NIGMS NIH HHS · GM27757 · United States
External Links
PubMed source DOI: 10.1017/s1355838299990027 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