F-Box Protein Specificity for G1 Cyclins Is Dictated by Subcellular Localization
Levels of G1 cyclins fluctuate in response to environmental cues and couple mitotic signaling to cell cycle entry. The G1 cyclin Cln3 is a key regulator of cell size and cell cycle entry in budding yeast. Cln3 degradation is essential for proper cell cycle control; however, the mechanisms that control Cln3 degradation are largely unknown. Here we show that two SCF ubiquitin ligases, SCFCdc4 and SCFGrr1, redundantly target Cln3 for degradation. While the F-box proteins (FBPs) Cdc4 and Grr1 were previously thought to target non-overlapping sets of substrates, we find that Cdc4 and Grr1 each bind to all 3 G1 cyclins in cell extracts, yet only Cln3 is redundantly targeted in vivo, due in part to its nuclear localization. The related cyclin Cln2 is cytoplasmic and exclusively targeted by Grr1. However, Cdc4 can interact with Cdk-phosphorylated Cln2 and target it for degradation when cytoplasmic Cdc4 localization is forced in vivo. These findings suggest that Cdc4 and Grr1 may share additional redundant targets and, consistent with this possibility, grr1Δ cdc4-1 cells demonstrate a CLN3-independent synergistic growth defect. Our findings demonstrate that structurally distinct FBPs are capable of interacting with some of the same substrates; however, in vivo specificity is achieved in part by subcellular localization. Additionally, the FBPs Cdc4 and Grr1 are partially redundant for proliferation and viability, likely sharing additional redundant substrates whose degradation is important for cell cycle progression.
Vyšlo v časopise:
F-Box Protein Specificity for G1 Cyclins Is Dictated by Subcellular Localization. PLoS Genet 8(7): e32767. doi:10.1371/journal.pgen.1002851
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002851
Souhrn
Levels of G1 cyclins fluctuate in response to environmental cues and couple mitotic signaling to cell cycle entry. The G1 cyclin Cln3 is a key regulator of cell size and cell cycle entry in budding yeast. Cln3 degradation is essential for proper cell cycle control; however, the mechanisms that control Cln3 degradation are largely unknown. Here we show that two SCF ubiquitin ligases, SCFCdc4 and SCFGrr1, redundantly target Cln3 for degradation. While the F-box proteins (FBPs) Cdc4 and Grr1 were previously thought to target non-overlapping sets of substrates, we find that Cdc4 and Grr1 each bind to all 3 G1 cyclins in cell extracts, yet only Cln3 is redundantly targeted in vivo, due in part to its nuclear localization. The related cyclin Cln2 is cytoplasmic and exclusively targeted by Grr1. However, Cdc4 can interact with Cdk-phosphorylated Cln2 and target it for degradation when cytoplasmic Cdc4 localization is forced in vivo. These findings suggest that Cdc4 and Grr1 may share additional redundant targets and, consistent with this possibility, grr1Δ cdc4-1 cells demonstrate a CLN3-independent synergistic growth defect. Our findings demonstrate that structurally distinct FBPs are capable of interacting with some of the same substrates; however, in vivo specificity is achieved in part by subcellular localization. Additionally, the FBPs Cdc4 and Grr1 are partially redundant for proliferation and viability, likely sharing additional redundant substrates whose degradation is important for cell cycle progression.
Zdroje
1. ReedSI 2003 Ratchets and clocks: the cell cycle, ubiquitylation and protein turnover. Nat Rev Mol Cell Biol 4 855 864
2. BaiCSenPHofmannKMaLGoeblM 1996 SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box. Cell 86 263 274
3. FeldmanRMCorrellCCKaplanKBDeshaiesRJ 1997 A complex of Cdc4p, Skp1p, and Cdc53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p. Cell 91 221 230
4. SkowyraDCraigKLTyersMElledgeSJHarperJW 1997 F-box proteins are receptors that recruit phosphorylated substrates to the SCF ubiquitin-ligase complex. Cell 91 209 219
5. SkowyraDKoeppDMKamuraTConradMNConawayRC 1999 Reconstitution of G1 cyclin ubiquitination with complexes containing SCFGrr1 and Rbx1. Science 284 662 665
6. OhtaTMichelJJSchotteliusAJXiongY 1999 ROC1, a homolog of APC11, represents a family of cullin partners with an associated ubiquitin ligase activity. Mol Cell 3 535 541
7. SeolJHFeldmanRMZachariaeWShevchenkoACorrellCC 1999 Cdc53/cullin and the essential Hrt1 RING-H2 subunit of SCF define a ubiquitin ligase module that activates the E2 enzyme Cdc34. Genes Dev 13 1614 1626
8. WillemsARSchwabMTyersM 2004 A hitchhiker's guide to the cullin ubiquitin ligases: SCF and its kin. Biochim Biophys Acta 1695 133 170
9. BarralYJentschSMannC 1995 G1 cyclin turnover and nutrient uptake are controlled by a common pathway in yeast. Genes Dev 9 399 409
10. VermaRFeldmanRMDeshaiesRJ 1997 SIC1 is ubiquitinated in vitro by a pathway that requires CDC4, CDC34, and cyclin/CDK activities. Mol Biol Cell 8 1427 1437
11. DruryLSPerkinsGDiffleyJF 1997 The Cdc4/34/53 pathway targets Cdc6p for proteolysis in budding yeast. EMBO J 16 5966 5976
12. HoltLJTuchBBVillenJJohnsonADGygiSP 2009 Global analysis of Cdk1 substrate phosphorylation sites provides insights into evolution. Science 325 1682 1686
13. UbersaxJAWoodburyELQuangPNParazMBlethrowJD 2003 Targets of the cyclin-dependent kinase Cdk1. Nature 425 859 864
14. BelleATanayABitinckaLShamirRO'SheaEK 2006 Quantification of protein half-lives in the budding yeast proteome. Proc Natl Acad Sci U S A 103 13004 13009
15. BenantiJACheungSKBradyMCToczyskiDP 2007 A proteomic screen reveals SCFGrr1 targets that regulate the glycolytic-gluconeogenic switch. Nat Cell Biol 9 1184 1191
16. TangXOrlickySLiuQWillemsASicheriF 2005 Genome-wide surveys for phosphorylation-dependent substrates of SCF ubiquitin ligases. Methods Enzymol 399 433 458
17. CostanzoMNishikawaJLTangXMillmanJSSchubO 2004 CDK activity antagonizes Whi5, an inhibitor of G1/S transcription in yeast. Cell 117 899 913
18. de BruinRAMcDonaldWHKalashnikovaTIYatesJ3rdWittenbergC 2004 Cln3 activates G1-specific transcription via phosphorylation of the SBF bound repressor Whi5. Cell 117 887 898
19. PramilaTWuWMilesSNobleWSBreedenLL 2006 The Forkhead transcription factor Hcm1 regulates chromosome segregation genes and fills the S-phase gap in the transcriptional circuitry of the cell cycle. Genes Dev 20 2266 2278
20. SpellmanPTSherlockGZhangMQIyerVRAndersK 1998 Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. Mol Biol Cell 9 3273 3297
21. EdgingtonNPFutcherB 2001 Relationship between the function and the location of G1 cyclins in S. cerevisiae. J Cell Sci 114 4599 4611
22. MillerMECrossFR 2001 Mechanisms controlling subcellular localization of the G(1) cyclins Cln2p and Cln3p in budding yeast. Mol Cell Biol 21 6292 6311
23. VergesEColominaNGariEGallegoCAldeaM 2007 Cyclin Cln3 is retained at the ER and released by the J chaperone Ydj1 in late G1 to trigger cell cycle entry. Mol Cell 26 649 662
24. CrossFR 1988 DAF1, a mutant gene affecting size control, pheromone arrest, and cell cycle kinetics of Saccharomyces cerevisiae. Mol Cell Biol 8 4675 4684
25. NashRTokiwaGAnandSEricksonKFutcherAB 1988 The WHI1+ gene of Saccharomyces cerevisiae tethers cell division to cell size and is a cyclin homolog. EMBO J 7 4335 4346
26. TyersMTokiwaGNashRFutcherB 1992 The Cln3-Cdc28 kinase complex of S. cerevisiae is regulated by proteolysis and phosphorylation. EMBO J 11 1773 1784
27. YaglomJLinskensMHSadisSRubinDMFutcherB 1995 p34Cdc28-mediated control of Cln3 cyclin degradation. Mol Cell Biol 15 731 741
28. MillerMECrossFR 2000 Distinct subcellular localization patterns contribute to functional specificity of the Cln2 and Cln3 cyclins of Saccharomyces cerevisiae. Mol Cell Biol 20 542 555
29. ChoRJCampbellMJWinzelerEASteinmetzLConwayA 1998 A genome-wide transcriptional analysis of the mitotic cell cycle. Mol Cell 2 65 73
30. HenchozSChiYCatarinBHerskowitzIDeshaiesRJ 1997 Phosphorylation- and ubiquitin-dependent degradation of the cyclin-dependent kinase inhibitor Far1p in budding yeast. Genes Dev 11 3046 3060
31. LyonsNAMorganDO 2011 Cdk1-dependent destruction of Eco1 prevents cohesion establishment after S phase. Mol Cell 42 378 389
32. KishiTYamaoF 1998 An essential function of Grr1 for the degradation of Cln2 is to act as a binding core that links Cln2 to Skp1. J Cell Sci 111 Pt 24 3655 3661
33. CrossFRBlakeCM 1993 The yeast Cln3 protein is an unstable activator of Cdc28. Mol Cell Biol 13 3266 3271
34. HaoBOehlmannSSowaMEHarperJWPavletichNP 2007 Structure of a Fbw7-Skp1-cyclin E complex: multisite-phosphorylated substrate recognition by SCF ubiquitin ligases. Mol Cell 26 131 143
35. KoivomagiMValkEVentaRIofikALepikuM 2011 Cascades of multisite phosphorylation control Sic1 destruction at the onset of S phase. Nature 480 128 131
36. SkaarJRPaganJKPaganoM 2009 SnapShot: F box proteins I. Cell 137 1160 1160 e1161
37. LankerSValdiviesoMHWittenbergC 1996 Rapid degradation of the G1 cyclin Cln2 induced by CDK-dependent phosphorylation. Science 271 1597 1601
38. BersetCGriacPTempelRLa RueJWittenbergC 2002 Transferable domain in the G(1) cyclin Cln2 sufficient to switch degradation of Sic1 from the E3 ubiquitin ligase SCF(Cdc4) to SCF(Grr1). Mol Cell Biol 22 4463 4476
39. BlondelMGalanJMChiYLafourcadeCLongarettiC 2000 Nuclear-specific degradation of Far1 is controlled by the localization of the F-box protein Cdc4. EMBO J 19 6085 6097
40. DixonSJCostanzoMBaryshnikovaAAndrewsBBooneC 2009 Systematic mapping of genetic interaction networks. Annu Rev Genet 43 601 625
41. HoltLJKrutchinskyANMorganDO 2008 Positive feedback sharpens the anaphase switch. Nature 454 353 357
42. StarostinaNGKipreosET 2012 Multiple degradation pathways regulate versatile CIP/KIP CDK inhibitors. Trends Cell Biol 22 33 41
43. XieYRubensteinEMMattTHochstrasserM 2010 SUMO-independent in vivo activity of a SUMO-targeted ubiquitin ligase toward a short-lived transcription factor. Genes Dev 24 893 903
44. DaiMSJinYGallegosJRLuH 2006 Balance of Yin and Yang: ubiquitylation-mediated regulation of p53 and c-Myc. Neoplasia 8 630 644
45. SkotheimJMDi TaliaSSiggiaEDCrossFR 2008 Positive feedback of G1 cyclins ensures coherent cell cycle entry. Nature 454 291 296
46. LinDIBarbashOKumarKGWeberJDHarperJW 2006 Phosphorylation-dependent ubiquitination of cyclin D1 by the SCF(FBX4-alphaB crystallin) complex. Mol Cell 24 355 366
47. KanieTOnoyamaIMatsumotoAYamadaMNakatsumiH 2011 Genetic reevaluation of the role of F-box proteins in cyclin D1 degradation. Mol Cell Biol
48. OkabeHLeeSHPhuchareonJAlbertsonDGMcCormickF 2006 A critical role for FBXW8 and MAPK in cyclin D1 degradation and cancer cell proliferation. PLoS ONE 1 e128 doi:10.1371/journal.pone.0000128
49. SantraMKWajapeyeeNGreenMR 2009 F-box protein FBXO31 mediates cyclin D1 degradation to induce G1 arrest after DNA damage. Nature 459 722 725
50. WeiSYangHCChuangHCYangJKulpSK 2008 A novel mechanism by which thiazolidinediones facilitate the proteasomal degradation of cyclin D1 in cancer cells. J Biol Chem 283 26759 26770
51. YuZKGervaisJLZhangH 1998 Human CUL-1 associates with the SKP1/SKP2 complex and regulates p21(CIP1/WAF1) and cyclin D proteins. Proc Natl Acad Sci U S A 95 11324 11329
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2012 Číslo 7
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