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PMID: 10712509 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Degradation of the transcription factor Gcn4 requires the kinase Pho85 and the SCF(CDC4) ubiquitin-ligase complex.

Molecular biology of the cell ·Vol. 11 ·No. 3 ·2000-03-00 ·Pages 915-27

Meimoun A, Holtzman T, Weissman Z, McBride HJ, Stillman DJ, Fink GR, Kornitzer D

Abstract

Gcn4, a yeast transcriptional activator that promotes the expression of amino acid and purine biosynthesis genes, is rapidly degraded in rich medium. Here we report that SCF(CDC4), a recently characterized protein complex that acts in conjunction with the ubiquitin-conjugating enzyme Cdc34 to degrade cell cycle regulators, is also necessary for the degradation of the transcription factor Gcn4. Degradation of Gcn4 occurs throughout the cell cycle, whereas degradation of the known cell cycle substrates of Cdc34/SCF(CDC4) is cell cycle regulated. Gcn4 ubiquitination and degradation are regulated by starvation for amino acids, whereas the degradation of the cell cycle substrates of Cdc34/SCF(CDC4) is unaffected by starvation. We further show that unlike the cell cycle substrates of Cdc34/SCF(CDC4), which require phosphorylation by the kinase Cdc28, Gcn4 degradation requires the kinase Pho85. We identify the critical target site of Pho85 on Gcn4; a mutation of this site stabilizes the protein. A specific Pho85-Pcl complex that is able to phosphorylate Gcn4 on that site is inactive under conditions under which Gcn4 is stable. Thus, Cdc34/SCF(CDC4) activity is constitutive, and regulation of the stability of its various substrates occurs at the level of their phosphorylation.

MeSH Terms
Cyclin-Dependent Kinases/metabolism DNA-Binding Proteins Enzyme Stability Fungal Proteins/metabolism Peptide Synthases/metabolism Phosphorylation Protein Biosynthesis Protein Kinases/metabolism SKP Cullin F-Box Protein Ligases Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins Signal Transduction Threonine/metabolism
Chemicals
DNA-Binding Proteins Fungal Proteins Saccharomyces cerevisiae Proteins Threonine SKP Cullin F-Box Protein Ligases Protein Kinases Cyclin-Dependent Kinases PHO85 protein, S cerevisiae Peptide Synthases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Meimoun A
Department of Microbiology, Technion-B. Rappaport Faculty of Medicine, Haifa 31096, Israel.
Holtzman T
Weissman Z
McBride H J
Stillman D J
Fink G R
Kornitzer D
References (66)
66 references, click to expand
  1. The Cdc4/34/53 pathway targets Cdc6p for proteolysis in budding yeast.
    EMBO J. 1997 Oct 1;16(19):5966-76 PMID: 9312054
  2. Coupling of cell division to cell growth by translational control of the G1 cyclin CLN3 in yeast.
    Genes Dev. 1997 Oct 1;11(19):2522-31 PMID: 9334317
  3. A kinase-cyclin pair in the RNA polymerase II holoenzyme.
    Nature. 1995 Mar 9;374(6518):193-6 PMID: 7877695
  4. G1 cyclin turnover and nutrient uptake are controlled by a common pathway in yeast.
    Genes Dev. 1995 Feb 15;9(4):399-409 PMID: 7883165
  5. A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B.
    Cell. 1995 Apr 21;81(2):279-88 PMID: 7736580
  6. A complex of Cdc4p, Skp1p, and Cdc53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p.
    Cell. 1997 Oct 17;91(2):221-30 PMID: 9346239
  7. Phosphorylation- and ubiquitin-dependent degradation of the cyclin-dependent kinase inhibitor Far1p in budding yeast.
    Genes Dev. 1997 Nov 15;11(22):3046-60 PMID: 9367986
  8. An essential domain within Cdc34p is required for binding to a complex containing Cdc4p and Cdc53p in Saccharomyces cerevisiae.
    J Biol Chem. 1998 Feb 13;273(7):4040-5 PMID: 9461595
  9. Cdc53 is a scaffold protein for multiple Cdc34/Skp1/F-box proteincomplexes that regulate cell division and methionine biosynthesis in yeast.
    Genes Dev. 1998 Mar 1;12(5):692-705 PMID: 9499404
  10. The cyclin family of budding yeast: abundant use of a good idea.
    Trends Genet. 1998 Feb;14(2):66-72 PMID: 9520600
  11. Cyclin partners determine Pho85 protein kinase substrate specificity in vitro and in vivo: control of glycogen biosynthesis by Pcl8 and Pcl10.
    Mol Cell Biol. 1998 Jun;18(6):3289-99 PMID: 9584169
  12. Combinatorial control in ubiquitin-dependent proteolysis: don't Skp the F-box hypothesis.
    Trends Genet. 1998 Jun;14(6):236-43 PMID: 9635407
  13. Cdc34 and the F-box protein Met30 are required for degradation of the Cdk-inhibitory kinase Swe1.
    Genes Dev. 1998 Aug 15;12(16):2587-97 PMID: 9716410
  14. Phosphorylation of sic1, a cyclin-dependent kinase (Cdk) inhibitor, by Cdk including Pho85 kinase is required for its prompt degradation.
    Mol Biol Cell. 1998 Sep;9(9):2393-405 PMID: 9725902
  15. The Cdc42p effector Gic2p is targeted for ubiquitin-dependent degradation by the SCFGrr1 complex.
    EMBO J. 1998 Sep 15;17(18):5360-73 PMID: 9736614
  16. The ubiquitin system.
    Annu Rev Biochem. 1998;67:425-79 PMID: 9759494
  17. A novel site for ubiquitination: the N-terminal residue, and not internal lysines of MyoD, is essential for conjugation and degradation of the protein.
    EMBO J. 1998 Oct 15;17(20):5964-73 PMID: 9774340
  18. Identification of the receptor component of the IkappaBalpha-ubiquitin ligase.
    Nature. 1998 Dec 10;396(6711):590-4 PMID: 9859996
  19. The SCFbeta-TRCP-ubiquitin ligase complex associates specifically with phosphorylated destruction motifs in IkappaBalpha and beta-catenin and stimulates IkappaBalpha ubiquitination in vitro.
    Genes Dev. 1999 Feb 1;13(3):270-83 PMID: 9990852
  20. Signal-induced ubiquitination of IkappaBalpha by the F-box protein Slimb/beta-TrCP.
    Genes Dev. 1999 Feb 1;13(3):284-94 PMID: 9990853
  21. The Cdc6 protein is ubiquitinated in vivo for proteolysis in Saccharomyces cerevisiae.
    J Biol Chem. 1999 Mar 26;274(13):9092-7 PMID: 10085159
  22. Rbx1, a component of the VHL tumor suppressor complex and SCF ubiquitin ligase.
    Science. 1999 Apr 23;284(5414):657-61 PMID: 10213691
  23. Reconstitution of G1 cyclin ubiquitination with complexes containing SCFGrr1 and Rbx1.
    Science. 1999 Apr 23;284(5414):662-5 PMID: 10213692
  24. CLN3 expression is sufficient to restore G1-to-S-phase progression in Saccharomyces cerevisiae mutants defective in translation initiation factor eIF4E.
    Biochem J. 1999 May 15;340 ( Pt 1):135-41 PMID: 10229668
  25. Roles of phosphorylation sites in regulating activity of the transcription factor Pho4.
    Science. 1999 May 7;284(5416):977-80 PMID: 10320381
  26. Substrate targeting of the yeast cyclin-dependent kinase Pho85p by the cyclin Pcl10p.
    Mol Cell Biol. 1999 Oct;19(10):7020-30 PMID: 10490639
  27. Phosphorylation controls timing of Cdc6p destruction: A biochemical analysis.
    Mol Biol Cell. 1999 Oct;10(10):3263-77 PMID: 10512865
  28. The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis.
    Mol Biol Cell. 1995 Feb;6(2):185-97 PMID: 7787245
  29. The histidyl-tRNA synthetase-related sequence in the eIF-2 alpha protein kinase GCN2 interacts with tRNA and is required for activation in response to starvation for different amino acids.
    Mol Cell Biol. 1995 Aug;15(8):4497-506 PMID: 7623840
  30. Regulation of PHO4 nuclear localization by the PHO80-PHO85 cyclin-CDK complex.
    Science. 1996 Jan 12;271(5246):209-12 PMID: 8539622
  31. Rapid degradation of the G1 cyclin Cln2 induced by CDK-dependent phosphorylation.
    Science. 1996 Mar 15;271(5255):1597-601 PMID: 8599119
  32. Linkage of replication to start by the Cdk inhibitor Sic1.
    Science. 1996 Apr 26;272(5261):560-2 PMID: 8614808
  33. SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box.
    Cell. 1996 Jul 26;86(2):263-74 PMID: 8706131
  34. Budding yeast SKP1 encodes an evolutionarily conserved kinetochore protein required for cell cycle progression.
    Cell. 1996 Jul 26;86(2):275-85 PMID: 8706132
  35. Cdc53 targets phosphorylated G1 cyclins for degradation by the ubiquitin proteolytic pathway.
    Cell. 1996 Aug 9;86(3):453-63 PMID: 8756727
  36. Phosphorylation of Sic1p by G1 Cdk required for its degradation and entry into S phase.
    Science. 1997 Oct 17;278(5337):455-60 PMID: 9334303
  37. F-box proteins are receptors that recruit phosphorylated substrates to the SCF ubiquitin-ligase complex.
    Cell. 1997 Oct 17;91(2):209-19 PMID: 9346238
  38. Control of cell division in Saccharomyces cerevisiae by methionyl-tRNA.
    Proc Natl Acad Sci U S A. 1976 May;73(5):1664-8 PMID: 775494
  39. Regulation of cell-cycle initiation in yeast by nutrients and protein synthesis.
    J Cell Physiol. 1978 Nov;97(2):177-87 PMID: 359576
  40. Positive regulation in the general amino acid control of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1983 Sep;80(17):5374-8 PMID: 6351059
  41. Copper metallothionein of yeast, structure of the gene, and regulation of expression.
    Proc Natl Acad Sci U S A. 1984 Jun;81(11):3332-6 PMID: 6374656
  42. Evidence for translational regulation of the activator of general amino acid control in yeast.
    Proc Natl Acad Sci U S A. 1984 Oct;81(20):6442-6 PMID: 6387704
  43. GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast.
    Cell. 1985 Nov;43(1):177-88 PMID: 3907851
  44. The relationship between the "TATA" sequence and transcription initiation sites at the HIS4 gene of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8557-61 PMID: 3909147
  45. Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast.
    Cell. 1986 Sep 12;46(6):885-94 PMID: 3530496
  46. Multiple global regulators control HIS4 transcription in yeast.
    Science. 1987 Aug 21;237(4817):874-80 PMID: 3303332
  47. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains.
    Genetics. 1987 Aug;116(4):541-5 PMID: 3305158
  48. Structural comparison of the yeast cell division cycle gene CDC4 and a related pseudogene.
    J Mol Biol. 1987 May 20;195(2):233-45 PMID: 3309335
  49. Structural and functional characterization of the short acidic transcriptional activation region of yeast GCN4 protein.
    Nature. 1988 Jun 16;333(6174):635-40 PMID: 3287180
  50. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  51. G1 events and regulation of cell proliferation.
    Science. 1989 Nov 3;246(4930):603-8 PMID: 2683075
  52. In vivo degradation of a transcriptional regulator: the yeast alpha 2 repressor.
    Cell. 1990 May 18;61(4):697-708 PMID: 2111732
  53. Phosphorylation of initiation factor 2 alpha by protein kinase GCN2 mediates gene-specific translational control of GCN4 in yeast.
    Cell. 1992 Feb 7;68(3):585-96 PMID: 1739968
  54. Coregulation of purine and histidine biosynthesis by the transcriptional activators BAS1 and BAS2.
    Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6746-50 PMID: 1495962
  55. The kin28 protein kinase is associated with a cyclin in Saccharomyces cerevisiae.
    J Mol Biol. 1993 Nov 20;234(2):307-10 PMID: 8230216
  56. Phosphorylation of the transcription factor PHO4 by a cyclin-CDK complex, PHO80-PHO85.
    Science. 1994 Feb 25;263(5150):1153-6 PMID: 8108735
  57. Cell cycle control by a complex of the cyclin HCS26 (PCL1) and the kinase PHO85.
    Science. 1994 Nov 25;266(5189):1388-91 PMID: 7973730
  58. The PCL2 (ORFD)-PHO85 cyclin-dependent kinase complex: a cell cycle regulator in yeast.
    Science. 1994 Nov 25;266(5189):1391-5 PMID: 7973731
  59. Regulated degradation of the transcription factor Gcn4.
    EMBO J. 1994 Dec 15;13(24):6021-30 PMID: 7813440
  60. The transcriptional activator GCN4 contains multiple activation domains that are critically dependent on hydrophobic amino acids.
    Mol Cell Biol. 1995 Mar;15(3):1220-33 PMID: 7862116
  61. The cyclin-dependent kinase inhibitor p40SIC1 imposes the requirement for Cln G1 cyclin function at Start.
    Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7772-6 PMID: 8755551
  62. Identification of seven hydrophobic clusters in GCN4 making redundant contributions to transcriptional activation.
    Mol Cell Biol. 1996 Oct;16(10):5557-71 PMID: 8816468
  63. The role of proteolysis in cell cycle progression in Schizosaccharomyces pombe.
    EMBO J. 1996 Oct 1;15(19):5268-79 PMID: 8895572
  64. Cdc53p acts in concert with Cdc4p and Cdc34p to control the G1-to-S-phase transition and identifies a conserved family of proteins.
    Mol Cell Biol. 1996 Dec;16(12):6634-43 PMID: 8943317
  65. A family of cyclin-like proteins that interact with the Pho85 cyclin-dependent kinase.
    Mol Cell Biol. 1997 Mar;17(3):1212-23 PMID: 9032248
  66. Translational regulation of yeast GCN4. A window on factors that control initiator-trna binding to the ribosome.
    J Biol Chem. 1997 Aug 29;272(35):21661-4 PMID: 9268289
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.11.3.915
Published
2000-03-00
Pages
915-27
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC14820
Subset
IM
Grants
NIGMS NIH HHS · GM39067 · United States
NIGMS NIH HHS · R01 GM039067 · United States
NIGMS NIH HHS · R01 GM048624 · United States
NIGMS NIH HHS · GM48624 · United States
NIGMS NIH HHS · R01 GM035010 · United States
NIGMS NIH HHS · T32 GM007464 · United States
NIGMS NIH HHS · GM35010 · United States
External Links
PubMed source DOI: 10.1091/mbc.11.3.915 PMC Full Text View journal details
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