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PMID: 15215893 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A complex containing the CCR4 and CAF1 proteins is involved in mRNA deadenylation in Drosophila.

The EMBO journal ·Vol. 23 ·No. 14 ·2004-07-21 ·Pages 2862-71

Temme C, Zaessinger S, Meyer S, Simonelig M, Wahle E

Abstract

The CCR4-NOT complex is the major enzyme catalyzing mRNA deadenylation in Saccharomyces cerevisiae. We have identified homologs for almost all subunits of this complex in the Drosophila genome. Biochemical fractionation showed that the two likely catalytic subunits, CCR4 and CAF1, were associated with each other and with a poly(A)-specific 3' exonuclease activity. In Drosophila, the CCR4 and CAF1 proteins were ubiquitously expressed and present in cytoplasmic foci. Individual knock-down of several potential subunits of the Drosophila CCR4-NOT complex by RNAi in tissue culture cells led to a lengthening of bulk mRNA poly(A) tails. Knock-down of two individual subunits also interfered with the rapid deadenylation of Hsp70 mRNA during recovery from heat shock. Similarly, ccr4 mutant flies had elongated bulk poly(A) and a defect in Hsp70 mRNA deadenylation. A minor increase in bulk poly(A) tail length was also observed in Rga mutant flies, which are affected in the NOT2 subunit. The data show that the CCR4-NOT complex is conserved in Drosophila melanogaster and plays a role in general and regulated mRNA deadenylation.

MeSH Terms
Amino Acid Sequence Animals Catalytic Domain Cells, Cultured Chromosomal Proteins, Non-Histone/genetics,metabolism Conserved Sequence Cytoplasm/metabolism Drosophila/cytology,metabolism Drosophila Proteins/genetics,metabolism HSP70 Heat-Shock Proteins/metabolism Molecular Chaperones/genetics,metabolism Molecular Sequence Data Mutation Protein Processing, Post-Translational Protein Structure, Tertiary RNA Interference RNA, Messenger/metabolism Retinoblastoma-Binding Protein 4 Ribonucleases/chemistry,genetics,metabolism Saccharomyces cerevisiae Proteins/chemistry,genetics Sequence Homology, Amino Acid
Chemicals
Caf1-55 protein, Drosophila Chromosomal Proteins, Non-Histone Drosophila Proteins HSP70 Heat-Shock Proteins Molecular Chaperones RNA, Messenger Retinoblastoma-Binding Protein 4 Saccharomyces cerevisiae Proteins CCR4 protein, Drosophila CCR4 protein, S cerevisiae Ribonucleases POP2 protein, S cerevisiae
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Temme Claudia
Institut für Biochemie, Martin-Luther-Universität Halle-Wittenberg, Halle, Germany.
Zaessinger Sophie
Meyer Sylke
Simonelig Martine
Wahle Elmar
References (51)
51 references, click to expand
  1. Functional organization of the yeast proteome by systematic analysis of protein complexes.
    Nature. 2002 Jan 10;415(6868):141-7 PMID: 11805826
  2. Preferential deadenylation of Hsp70 mRNA plays a key role in regulating Hsp70 expression in Drosophila melanogaster.
    Mol Cell Biol. 1994 Jun;14(6):3646-59 PMID: 7515148
  3. Isolation and characterization of human orthologs of yeast CCR4-NOT complex subunits.
    Nucleic Acids Res. 2000 Feb 1;28(3):809-17 PMID: 10637334
  4. 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
  5. The Drosophila poly(A) binding protein-interacting protein, dPaip2, is a novel effector of cell growth.
    Mol Cell Biol. 2004 Feb;24(3):1143-54 PMID: 14729960
  6. Regena (Rga), a Drosophila homolog of the global negative transcriptional regulator CDC36 (NOT2) from yeast, modifies gene expression and suppresses position effect variegation.
    Genetics. 1998 Jan;148(1):317-29 PMID: 9475742
  7. Stimulation of poly(A) polymerase through a direct interaction with the nuclear poly(A) binding protein allosterically regulated by RNA.
    EMBO J. 2003 Jul 15;22(14):3705-14 PMID: 12853485
  8. Global control of gene expression in yeast by the Ccr4-Not complex.
    Gene. 2003 Aug 14;313:1-16 PMID: 12957374
  9. mRNA stability in mammalian cells.
    Microbiol Rev. 1995 Sep;59(3):423-50 PMID: 7565413
  10. Saccharomyces cerevisiae Ccr4-not complex contributes to the control of Msn2p-dependent transcription by the Ras/cAMP pathway.
    Mol Microbiol. 2002 Feb;43(4):1023-37 PMID: 11929548
  11. Human Dcp2: a catalytically active mRNA decapping enzyme located in specific cytoplasmic structures.
    EMBO J. 2002 Dec 16;21(24):6915-24 PMID: 12486012
  12. Characterization of NOT5 that encodes a new component of the Not protein complex.
    Gene. 1998 Jan 19;207(1):61-9 PMID: 9511744
  13. The CCR4 and CAF1 proteins of the CCR4-NOT complex are physically and functionally separated from NOT2, NOT4, and NOT5.
    Mol Cell Biol. 1999 Oct;19(10):6642-51 PMID: 10490603
  14. Identification of a human cytoplasmic poly(A) nuclease complex stimulated by poly(A)-binding protein.
    J Biol Chem. 2004 Jan 9;279(2):1383-91 PMID: 14583602
  15. CCR4, a 3'-5' poly(A) RNA and ssDNA exonuclease, is the catalytic component of the cytoplasmic deadenylase.
    EMBO J. 2002 Mar 15;21(6):1414-26 PMID: 11889047
  16. Cytoplasmic foci are sites of mRNA decay in human cells.
    J Cell Biol. 2004 Apr12;165(1):31-40 PMID: 15067023
  17. The Drosophila hsp70 message is rapidly degraded at normal temperatures and stabilized by heat shock.
    Gene. 1988 Dec 10;72(1-2):161-8 PMID: 3243430
  18. The CCR4-NOT complex plays diverse roles in mRNA metabolism.
    Prog Nucleic Acid Res Mol Biol. 2003;73:221-50 PMID: 12882519
  19. 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
  20. The enzymes and control of eukaryotic mRNA turnover.
    Nat Struct Mol Biol. 2004 Feb;11(2):121-7 PMID: 14749774
  21. DBF2, a cell cycle-regulated protein kinase, is physically and functionally associated with the CCR4 transcriptional regulatory complex.
    EMBO J. 1997 Sep 1;16(17):5289-98 PMID: 9311989
  22. Systematic mutagenesis of the leucine-rich repeat (LRR) domain of CCR4 reveals specific sites for binding to CAF1 and a separate critical role for the LRR in CCR4 deadenylase activity.
    J Biol Chem. 2004 Apr 2;279(14):13616-23 PMID: 14734555
  23. Yak1p, a DYRK family kinase, translocates to the nucleus and phosphorylates yeast Pop2p in response to a glucose signal.
    Genes Dev. 2001 May 15;15(10):1217-28 PMID: 11358866
  24. The heat shock response in HeLa cells is accompanied by elevated expression of the c-fos proto-oncogene.
    Mol Cell Biol. 1987 Oct;7(10):3452-8 PMID: 3316977
  25. Effect of heat shock on RNA metabolism in HeLa cells.
    J Cell Physiol. 1988 Jun;135(3):377-86 PMID: 2456286
  26. mRNA decay enzymes: decappers conserved between yeast and mammals.
    Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12512-4 PMID: 12271148
  27. 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
  28. Purification and characterization of the 1.0 MDa CCR4-NOT complex identifies two novel components of the complex.
    J Mol Biol. 2001 Dec 7;314(4):683-94 PMID: 11733989
  29. Mutational analysis of mammalian poly(A) polymerase identifies a region for primer binding and catalytic domain, homologous to the family X polymerases, and to other nucleotidyltransferases.
    EMBO J. 1996 May 15;15(10):2593-603 PMID: 8665867
  30. Double-stranded RNA-dependent protein kinase (pkr) is essential for thermotolerance, accumulation of HSP70, and stabilization of ARE-containing HSP70 mRNA during stress.
    J Biol Chem. 2002 Nov 15;277(46):44539-47 PMID: 12207033
  31. Poly(A) tail shortening by a mammalian poly(A)-specific 3'-exoribonuclease.
    J Biol Chem. 1997 Apr 18;272(16):10448-56 PMID: 9099687
  32. The human LSm1-7 proteins colocalize with the mRNA-degrading enzymes Dcp1/2 and Xrnl in distinct cytoplasmic foci.
    RNA. 2002 Dec;8(12):1489-501 PMID: 12515382
  33. Identification of four families of yCCR4- and Mg2+-dependent endonuclease-related proteins in higher eukaryotes, and characterization of orthologs of yCCR4 with a conserved leucine-rich repeat essential for hCAF1/hPOP2 binding.
    BMC Genomics. 2001;2:9 PMID: 11747467
  34. AU-rich elements: characterization and importance in mRNA degradation.
    Trends Biochem Sci. 1995 Nov;20(11):465-70 PMID: 8578590
  35. Analysis of poly(A) tail lengths by PCR: the PAT assay.
    Methods Mol Biol. 1999;118:441-8 PMID: 10549542
  36. Ccr4p is the catalytic subunit of a Ccr4p/Pop2p/Notp mRNA deadenylase complex in Saccharomyces cerevisiae.
    EMBO J. 2002 Mar 15;21(6):1427-36 PMID: 11889048
  37. Targeting an mRNA for decapping: displacement of translation factors and association of the Lsm1p-7p complex on deadenylated yeast mRNAs.
    Mol Cell. 2001 Nov;8(5):1075-83 PMID: 11741542
  38. Messenger RNA deadenylylation precedes decapping in mammalian cells.
    Proc Natl Acad Sci U S A. 1997 May 27;94(11):5628-33 PMID: 9159123
  39. X-ray structure and activity of the yeast Pop2 protein: a nuclease subunit of the mRNA deadenylase complex.
    EMBO Rep. 2003 Dec;4(12):1150-5 PMID: 14618157
  40. 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
  41. 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
  42. Poly(A) tail length control in Saccharomyces cerevisiae occurs by message-specific deadenylation.
    Mol Cell Biol. 1998 Nov;18(11):6548-59 PMID: 9774670
  43. Nocturnin, a deadenylase in Xenopus laevis retina: a mechanism for posttranscriptional control of circadian-related mRNA.
    Curr Biol. 2003 Feb 4;13(3):189-98 PMID: 12573214
  44. The Drosophila poly(A)-binding protein II is ubiquitous throughout Drosophila development and has the same function in mRNA polyadenylation as its bovine homolog in vitro.
    Nucleic Acids Res. 1999 Oct 1;27(19):3771-8 PMID: 10481015
  45. Decapping and decay of messenger RNA occur in cytoplasmic processing bodies.
    Science. 2003 May 2;300(5620):805-8 PMID: 12730603
  46. The GW182 protein colocalizes with mRNA degradation associated proteins hDcp1 and hLSm4 in cytoplasmic GW bodies.
    RNA. 2003 Oct;9(10):1171-3 PMID: 13130130
  47. A novel embryonic poly(A) binding protein, ePAB, regulates mRNA deadenylation in Xenopus egg extracts.
    Genes Dev. 2001 Mar 15;15(6):774-88 PMID: 11274061
  48. Messenger RNA degradation: beginning at the end.
    Curr Biol. 2002 Apr 16;12(8):R285-7 PMID: 11967169
  49. The yeast POP2 gene encodes a nuclease involved in mRNA deadenylation.
    Nucleic Acids Res. 2001 Jun 15;29(12):2448-55 PMID: 11410650
  50. Control of mRNA decay by heat shock-ubiquitin-proteasome pathway.
    Science. 1999 Apr 16;284(5413):499-502 PMID: 10205060
  51. Regulation of HSP70 synthesis by messenger RNA degradation.
    Cell Regul. 1989 Nov;1(1):135-49 PMID: 2519613
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600273
Published
2004-07-21
Epub
2004-00-24
Pages
2862-71
Language
English
Region
England
NLM ID
8208664
PMCID
PMC514940
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.emboj.7600273 PMC Full Text View journal details
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