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PMID: 11410650 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The yeast POP2 gene encodes a nuclease involved in mRNA deadenylation.

Nucleic acids research ·Vol. 29 ·No. 12 ·2001-06-15 ·Pages 2448-55

Daugeron MC, Mauxion F, Séraphin B

Abstract

The major mRNA degradation pathway involves deadenylation of the target molecule followed by decapping and, finally, 5'-->3' exonuclease digestion of the mRNA body. While yeast factors involved in the decapping and exonuclease degradation steps have been identified, the nature of the factor(s) involved in the deadenylation step remained elusive. Database searches for yeast proteins related to the mammalian deadenylase PARN identified the Pop2 protein (Pop2p) as a potential deadenylase. While Pop2p was previously identified as a factor affecting transcription, we identified a non-canonical RNase D sequence signature in its sequence. Analysis of the fate of a reporter mRNA in a pop2 mutant demonstrates that Pop2p is required for efficient mRNA degradation in vivo. Characterisation of mRNA degradation intermediates accumulating in this mutant supports the involvement of Pop2p in mRNA deadenylation in vivo. Similar phenotypes are observed in yeast strains lacking the Ccr4 protein, which is known to be associated with Pop2p. A recombinant Pop2p fragment encompassing the putative catalytic domain degrades poly(A) in vitro demonstrating that Pop2p is a nuclease. We also demonstrate that poly(A) is a better competitor than poly(G) or poly(C) of the Pop2p nuclease activity. Altogether, our study indicates that Pop2p is a nuclease subunit of the yeast deadenylase and suggests that Pop2p homologues in other species may have similar functions.

MeSH Terms
Amino Acid Sequence Animals Catalytic Domain Computational Biology Endoribonucleases/chemistry,genetics,metabolism Fungal Proteins/genetics,metabolism Gene Deletion Genes, Fungal/genetics Humans Molecular Sequence Data Poly A/genetics Protein Structure, Tertiary Protein Subunits Proteins RNA Stability RNA, Fungal/chemistry,genetics,metabolism RNA, Messenger/chemistry,genetics,metabolism Ribonuclease H/metabolism Ribonuclease III Ribonucleases Saccharomyces cerevisiae Proteins Substrate Specificity Transcription Factors/chemistry,genetics,metabolism Yeasts/enzymology,genetics
Chemicals
CNOT8 protein, human Fungal Proteins Protein Subunits Proteins RNA, Fungal RNA, Messenger Saccharomyces cerevisiae Proteins Transcription Factors Poly A CCR4 protein, S cerevisiae Endoribonucleases Ribonucleases POP2 protein, S cerevisiae Ribonuclease III Ribonuclease H
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Daugeron M C
Centre de Génétique Moléculaire, CNRS, Avenue de la Terrasse, F-91198 Gif sur Yvette, France.
Mauxion F
Séraphin B
References (45)
45 references, click to expand
  1. The exosome: a proteasome for RNA?
    Cell. 1999 Nov 12;99(4):347-50 PMID: 10571176
  2. 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
  3. A Sm-like protein complex that participates in mRNA degradation.
    EMBO J. 2000 Apr 3;19(7):1661-71 PMID: 10747033
  4. Yeast Sm-like proteins function in mRNA decapping and decay.
    Nature. 2000 Mar 30;404(6777):515-8 PMID: 10761922
  5. Functionally unrelated signalling proteins contain a fold similar to Mg2+-dependent endonucleases.
    Trends Biochem Sci. 2000 Jun;25(6):272-3 PMID: 10838565
  6. The yeast hnRNP-like protein Hrp1/Nab4 marks a transcript for nonsense-mediated mRNA decay.
    Mol Cell. 2000 Mar;5(3):489-99 PMID: 10882134
  7. Genome-wide protein interaction screens reveal functional networks involving Sm-like proteins.
    Yeast. 2000 Jun 30;17(2):95-110 PMID: 10900456
  8. The two proteins Pat1p (Mrt1p) and Spb8p interact in vivo, are required for mRNA decay, and are functionally linked to Pab1p.
    Mol Cell Biol. 2000 Aug;20(16):5939-46 PMID: 10913177
  9. The eukaryotic mRNA decapping protein Dcp1 interacts physically and functionally with the eIF4F translation initiation complex.
    EMBO J. 2000 Aug 15;19(16):4372-82 PMID: 10944120
  10. Mechanisms and control of mRNA decapping in Saccharomyces cerevisiae.
    Annu Rev Biochem. 2000;69:571-95 PMID: 10966469
  11. The essential function of Not1 lies within the Ccr4-Not complex.
    J Mol Biol. 2000 Oct 20;303(2):131-43 PMID: 11023781
  12. Identification of a regulated pathway for nuclear pre-mRNA turnover.
    Cell. 2000 Sep 15;102(6):765-75 PMID: 11030620
  13. The Puf3 protein is a transcript-specific regulator of mRNA degradation in yeast.
    EMBO J. 2000 Dec 1;19(23):6602-11 PMID: 11101532
  14. Two related proteins, Edc1p and Edc2p, stimulate mRNA decapping in Saccharomyces cerevisiae.
    Genetics. 2001 Jan;157(1):27-37 PMID: 11139489
  15. Post-transcriptional regulation through the HO 3'-UTR by Mpt5, a yeast homolog of Pumilio and FBF.
    EMBO J. 2001 Feb 1;20(3):552-61 PMID: 11157761
  16. Exoribonuclease superfamilies: structural analysis and phylogenetic distribution.
    Nucleic Acids Res. 2001 Mar 1;29(5):1017-26 PMID: 11222749
  17. The transcription factor associated Ccr4 and Caf1 proteins are components of the major cytoplasmic mRNA deadenylase in Saccharomyces cerevisiae.
    Cell. 2001 Feb 9;104(3):377-86 PMID: 11239395
  18. 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
  19. Identification of a mouse protein whose homolog in Saccharomyces cerevisiae is a component of the CCR4 transcriptional regulatory complex.
    Mol Cell Biol. 1995 Jul;15(7):3487-95 PMID: 7791755
  20. The yeast Pan2 protein is required for poly(A)-binding protein-stimulated poly(A)-nuclease activity.
    J Biol Chem. 1996 Jan 5;271(1):432-8 PMID: 8550599
  21. An essential component of the decapping enzyme required for normal rates of mRNA turnover.
    Nature. 1996 Aug 15;382(6592):642-6 PMID: 8757137
  22. PAN3 encodes a subunit of the Pab1p-dependent poly(A) nuclease in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Oct;16(10):5744-53 PMID: 8816488
  23. Mechanisms and control of mRNA turnover in Saccharomyces cerevisiae.
    Microbiol Rev. 1996 Mar;60(1):233-49 PMID: 8852902
  24. Poly(A) tail shortening by a mammalian poly(A)-specific 3'-exoribonuclease.
    J Biol Chem. 1997 Apr 18;272(16):10448-56 PMID: 9099687
  25. Messenger RNA deadenylylation precedes decapping in mammalian cells.
    Proc Natl Acad Sci U S A. 1997 May 27;94(11):5628-33 PMID: 9159123
  26. 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
  27. Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae.
    Yeast. 1997 Sep 15;13(11):1065-75 PMID: 9290211
  28. Comparative sequence analysis of ribonucleases HII, III, II PH and D.
    Nucleic Acids Res. 1997 Aug 15;25(16):3187-95 PMID: 9241229
  29. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.
    Nucleic Acids Res. 1997 Dec 15;25(24):4876-82 PMID: 9396791
  30. The proofreading domain of Escherichia coli DNA polymerase I and other DNA and/or RNA exonuclease domains.
    Nucleic Acids Res. 1997 Dec 15;25(24):5110-8 PMID: 9396823
  31. The NOT proteins are part of the CCR4 transcriptional complex and affect gene expression both positively and negatively.
    EMBO J. 1998 Feb 16;17(4):1096-106 PMID: 9463387
  32. Capped mRNA degradation intermediates accumulate in the yeast spb8-2 mutant.
    Mol Cell Biol. 1998 Sep;18(9):5062-72 PMID: 9710590
  33. 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
  34. Poly(A) tail length control in Saccharomyces cerevisiae occurs by message-specific deadenylation.
    Mol Cell Biol. 1998 Nov;18(11):6548-59 PMID: 9774670
  35. Should we kill the messenger? The role of the surveillance complex in translation termination and mRNA turnover.
    Bioessays. 1999 Aug;21(8):685-96 PMID: 10440865
  36. The DCP2 protein is required for mRNA decapping in Saccharomyces cerevisiae and contains a functional MutT motif.
    EMBO J. 1999 Oct 1;18(19):5411-22 PMID: 10508173
  37. Mouse CAF1, a mouse homologue of the yeast POP2 gene, complements the yeast pop2 null mutation.
    Yeast. 1999 Sep 30;15(13):1357-64 PMID: 10509017
  38. Transfer RNA is a substrate for RNase D in vivo.
    J Biol Chem. 1988 Dec 5;263(34):17909-12 PMID: 3056931
  39. Elevated recombination rates in transcriptionally active DNA.
    Cell. 1989 Feb 24;56(4):619-30 PMID: 2645056
  40. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  41. Characterization of the XRN1 gene encoding a 5'-->3' exoribonuclease: sequence data and analysis of disparate protein and mRNA levels of gene-disrupted yeast cells.
    Gene. 1992 Oct 12;120(1):51-7 PMID: 1398123
  42. The CCR4 protein from Saccharomyces cerevisiae contains a leucine-rich repeat region which is required for its control of ADH2 gene expression.
    Genetics. 1992 Dec;132(4):951-62 PMID: 1459446
  43. 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
  44. 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
  45. Three conserved members of the RNase D family have unique and overlapping functions in the processing of 5S, 5.8S, U4, U5, RNase MRP and RNase P RNAs in yeast.
    EMBO J. 2000 Mar 15;19(6):1357-65 PMID: 10716935
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
Published
2001-06-15
Pages
2448-55
Language
English
Region
England
NLM ID
0411011
PMCID
PMC55743
Subset
IM
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