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PMID: 17452450 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Depletion of mammalian CCR4b deadenylase triggers elevation of the p27Kip1 mRNA level and impairs cell growth.

Molecular and cellular biology ·Vol. 27 ·No. 13 ·2007-07-00 ·Pages 4980-90

Morita M, Suzuki T, Nakamura T, Yokoyama K, Miyasaka T, Yamamoto T

Abstract

The stability of mRNA influences the abundance of cellular transcripts and proteins. Deadenylases play critical roles in mRNA turnover and thus are important for the regulation of various biological events. Here, we report the identification and characterization of CCR4b/CNOT6L, which is homologous to yeast CCR4 mRNA deadenylase. CCR4b is localized mainly in the cytoplasm and displays deadenylase activity both in vitro and in vivo. CCR4b forms a multisubunit complex similar to the yeast CCR4-NOT complex. Suppression of CCR4b by RNA interference results in growth retardation of NIH 3T3 cells accompanied by elevation of both p27(Kip1) mRNA and p27(Kip1) protein. Reintroduction of wild-type CCR4b, but not mutant CCR4b lacking deadenylase activity, restores the growth of CCR4b-depleted NIH 3T3 cells. The data suggest that CCR4b regulates cell growth in a manner dependent on its deadenylase activity. We also show that p27(Kip1) mRNA is stabilized and its poly(A) tail is preserved in CCR4b-depleted cells. Our findings provide evidence that CCR4b deadenylase is a constituent of the mammalian CCR4-NOT complex and regulates the turnover rate of specific target mRNAs. Thus, CCR4b may be involved in various cellular events that include cell proliferation.

MeSH Terms
Animals Cell Cycle Cell Line Cell Proliferation Cloning, Molecular Cyclin-Dependent Kinase Inhibitor p27/genetics,metabolism DNA, Complementary Exoribonucleases/metabolism Gene Expression Regulation Humans Mice NIH 3T3 Cells Poly A/metabolism Protein Binding Protein Transport RNA Stability RNA, Messenger/genetics,metabolism Ribonucleases/chemistry,deficiency Subcellular Fractions/metabolism Tissue Distribution
Chemicals
DNA, Complementary RNA, Messenger Cyclin-Dependent Kinase Inhibitor p27 Poly A CNOT6L protein, human Cnot6l protein, mouse Exoribonucleases Ribonucleases mRNA deadenylase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Morita Masahiro
Division of Oncology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan.
Suzuki Toru
Nakamura Takahisa
Yokoyama Kazumasa
Miyasaka Takashi
Yamamoto Tadashi
References (36)
36 references, click to expand
  1. Oligo-astheno-teratozoospermia in mice lacking Cnot7, a regulator of retinoid X receptor beta.
    Nat Genet. 2004 May;36(5):528-33 PMID: 15107851
  2. The ubiquitous transcription factor NF-Y positively regulates the transcription of human p27Kip1 through a CCAAT box located in the 5-upstream region of the p27Kip1 gene.
    FEBS Lett. 1999 Jul 23;455(3):281-5 PMID: 10437789
  3. Negative regulation of BMP/Smad signaling by Tob in osteoblasts.
    Cell. 2000 Dec 22;103(7):1085-97 PMID: 11163184
  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. A novel mRNA-decapping activity in HeLa cytoplasmic extracts is regulated by AU-rich elements.
    EMBO J. 2001 Mar 1;20(5):1134-43 PMID: 11230136
  6. The cap-to-tail guide to mRNA turnover.
    Nat Rev Mol Cell Biol. 2001 Apr;2(4):237-46 PMID: 11283721
  7. 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
  8. 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
  9. The telomeric poly(ADP-ribose) polymerase, tankyrase 1, contains multiple binding sites for telomeric repeat binding factor 1 (TRF1) and a novel acceptor, 182-kDa tankyrase-binding protein (TAB182).
    J Biol Chem. 2002 Apr 19;277(16):14116-26 PMID: 11854288
  10. Phosphorylation of three regulatory serines of Tob by Erk1 and Erk2 is required for Ras-mediated cell proliferation and transformation.
    Genes Dev. 2002 Jun 1;16(11):1356-70 PMID: 12050114
  11. Cell cycle-dependent translation of p27 involves a responsive element in its 5'-UTR that overlaps with a uORF.
    Hum Mol Genet. 2003 Jul 15;12(14):1767-79 PMID: 12837699
  12. p250GAP, a novel brain-enriched GTPase-activating protein for Rho family GTPases, is involved in the N-methyl-d-aspartate receptor signaling.
    Mol Biol Cell. 2003 Jul;14(7):2921-34 PMID: 12857875
  13. The CCR4-NOT complex plays diverse roles in mRNA metabolism.
    Prog Nucleic Acid Res Mol Biol. 2003;73:221-50 PMID: 12882519
  14. Global control of gene expression in yeast by the Ccr4-Not complex.
    Gene. 2003 Aug 14;313:1-16 PMID: 12957374
  15. The enzymes and control of eukaryotic mRNA turnover.
    Nat Struct Mol Biol. 2004 Feb;11(2):121-7 PMID: 14749774
  16. Cell cycle progression in G1 and S phases is CCR4 dependent following ionizing radiation or replication stress in Saccharomyces cerevisiae.
    Eukaryot Cell. 2004 Apr;3(2):430-46 PMID: 15075273
  17. Mouse CAF1 can function as a processive deadenylase/3'-5'-exonuclease in vitro but in yeast the deadenylase function of CAF1 is not required for mRNA poly(A) removal.
    J Biol Chem. 2004 Jun 4;279(23):23988-95 PMID: 15044470
  18. CCR4-associated factor CAF1 is an essential factor for spermatogenesis.
    Mol Cell Biol. 2004 Jul;24(13):5808-20 PMID: 15199137
  19. A complex containing the CCR4 and CAF1 proteins is involved in mRNA deadenylation in Drosophila.
    EMBO J. 2004 Jul 21;23(14):2862-71 PMID: 15215893
  20. UNR, a new partner of poly(A)-binding protein, plays a key role in translationally coupled mRNA turnover mediated by the c-fos major coding-region determinant.
    Genes Dev. 2004 Aug 15;18(16):2010-23 PMID: 15314026
  21. Nuclear localization of Tob is important for regulation of its antiproliferative activity.
    Oncogene. 2004 Aug 26;23(39):6630-8 PMID: 15235587
  22. Starting at the beginning, middle, and end: translation initiation in eukaryotes.
    Cell. 1997 Jun 13;89(6):831-8 PMID: 9200601
  23. 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
  24. Ccr4-not complex mRNA deadenylase activity contributes to DNA damage responses in Saccharomyces cerevisiae.
    Genetics. 2005 Jan;169(1):65-75 PMID: 15466434
  25. twin, a CCR4 homolog, regulates cyclin poly(A) tail length to permit Drosophila oogenesis.
    Development. 2005 Mar;132(6):1165-74 PMID: 15703281
  26. The yeast EDC1 mRNA undergoes deadenylation-independent decapping stimulated by Not2p, Not4p, and Not5p.
    EMBO J. 2005 Mar 9;24(5):1033-45 PMID: 15706350
  27. PDGF-BB regulates p27 expression through ERK-dependent RNA turn-over in vascular smooth muscle cells.
    J Biol Chem. 2005 Jul 8;280(27):25470-7 PMID: 15894805
  28. DNA damage and replication stress induced transcription of RNR genes is dependent on the Ccr4-Not complex.
    Nucleic Acids Res. 2005;33(19):6384-92 PMID: 16275785
  29. Concerted action of poly(A) nucleases and decapping enzyme in mammalian mRNA turnover.
    Nat Struct Mol Biol. 2005 Dec;12(12):1054-63 PMID: 16284618
  30. Oskar allows nanos mRNA translation in Drosophila embryos by preventing its deadenylation by Smaug/CCR4.
    Development. 2006 Nov;133(22):4573-83 PMID: 17050620
  31. Stabilization of interleukin-2 mRNA by the c-Jun NH2-terminal kinase pathway.
    Science. 1998 Jun 19;280(5371):1945-9 PMID: 9632395
  32. Poly(A) tail length control in Saccharomyces cerevisiae occurs by message-specific deadenylation.
    Mol Cell Biol. 1998 Nov;18(11):6548-59 PMID: 9774670
  33. Assaying the polyadenylation state of mRNAs.
    Methods. 1999 Jan;17(1):38-45 PMID: 10075881
  34. Evidence that tristetraprolin binds to AU-rich elements and promotes the deadenylation and destabilization of tumor necrosis factor alpha mRNA.
    Mol Cell Biol. 1999 Jun;19(6):4311-23 PMID: 10330172
  35. CDK inhibitors: positive and negative regulators of G1-phase progression.
    Genes Dev. 1999 Jun 15;13(12):1501-12 PMID: 10385618
  36. A mechanism for translationally coupled mRNA turnover: interaction between the poly(A) tail and a c-fos RNA coding determinant via a protein complex.
    Cell. 2000 Sep 29;103(1):29-40 PMID: 11051545
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.02304-06
Published
2007-07-00
Epub
2007-00-23
Pages
4980-90
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1951489
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.02304-06 PMC Full Text View journal details
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