Accumulation of a Threonine Biosynthetic Intermediate Attenuates General Amino Acid Control by Accelerating Degradation of Gcn4 via Pho85 and Cdk8

English version České info

Transcriptional activator Gcn4 maintains amino acid homeostasis in budding yeast by inducing multiple amino acid biosynthetic pathways in response to starvation for any amino acid—the general amino acid control. Gcn4 abundance is tightly regulated by the interplay between an intricate translational control mechanism, which induces Gcn4 synthesis in starved cells, and a pathway of phosphorylation and ubiquitylation that mediates its rapid degradation by the proteasome. Here, we discovered that accumulation of a threonine biosynthetic pathway intermediate, β-aspartate semialdehyde (ASA), in hom6Δ mutant cells impairs general amino acid control in cells starved for isoleucine and valine by accelerating the already rapid degradation of Gcn4, in a manner requiring its phosphorylation by cyclin-dependent kinases Cdk8/Srb10 and Pho85. Interestingly, our results unveil a division of labor between these two kinases wherein Srb10 primarily targets inactive Gcn4 molecules—presumably damaged under conditions of ASA excess—while Pho85 clears a greater proportion of functional Gcn4 species from the cell. The ability of ASA to inhibit transcriptional induction of threonine pathway enzymes by Gcn4, dampening ASA accumulation and its toxic effects on cell physiology, should be adaptive in the wild when yeast encounters natural antibiotics that target Hom6 enzymatic activity.

Vyšlo v časopise: Accumulation of a Threonine Biosynthetic Intermediate Attenuates General Amino Acid Control by Accelerating Degradation of Gcn4 via Pho85 and Cdk8. PLoS Genet 10(7): e32767. doi:10.1371/journal.pgen.1004534
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004534

Souhrn

Zdroje

1. Johnston M, Carlson M (1992) Regulation of carbon and phosphate utilization. In: Jones EW, Pringle JR, Broach JR, editors. The Molecular and Cellular Biology of the Yeast Saccharomyces: Gene Expression. Cold Spring Harbor: Cold Spring Harbor Press. pp. 193–282.

2. KaffmanA, O'SheaEK (1999) Regulation of nuclear localization: a key to a door. Annu Rev Cell Dev Biol 15 : 291–339.

3. HinnebuschAG (2005) Translational regulation of GCN4 and the general amino acid control of yeast. Annu Rev Microbiol 59 : 407–450.

4. JiaMH, LarossaRA, LeeJM, RafalskiA, DeroseE, et al. (2000) Global expression profiling of yeast treated with an inhibitor of amino acid biosynthesis, sulfometuron methyl. Physiol Genomics 3 : 83–92.

5. NatarajanK, MeyerMR, JacksonBM, SladeD, RobertsC, et al. (2001) Transcriptional profiling shows that Gcn4p is a master regulator of gene expression during amino acid starvation in yeast. Mol Cell Biol 21 : 4347–4368.

6. PriesR, BomekeK, IrnigerS, GrundmannO, BrausGH (2002) Amino acid-dependent Gcn4p stability regulation occurs exclusively in the yeast nucleus. Eukaryot Cell 1 : 663–672.

7. SwansonMJ, QiuH, SumibcayL, KruegerA, KimS-J, et al. (2003) A Multiplicity of coactivators is required by Gcn4p at individual promoters in vivo. MolCellBiol 23 : 2800–2820.

8. QiuH, HuC, ZhangF, HwangGJ, SwansonMJ, et al. (2005) Interdependent recruitment of SAGA and Srb mediator by transcriptional activator Gcn4p. Mol Cell Biol 25 : 3461–3474.

9. GovindCK, YoonS, QiuH, GovindS, HinnebuschAG (2005) Simultaneous recruitment of coactivators by Gcn4p stimulates multiple steps of transcription in vivo. Mol Cell Biol 25 : 5626–5638.

10. QiuH, HuC, YoonS, NatarajanK, SwansonMJ, et al. (2004) An array of coactivators is required for optimal recruitment of TATA binding protein and RNA polymerase II by promoter-bound Gcn4p. Mol Cell Biol 24 : 4104–4117.

11. KornitzerD, RaboyB, KulkaRG, FinkGR (1994) Regulated degradation of the transcription factor Gcn4. EMBO J 13 : 6021–6030.

12. MeimounA, HoltzmanT, WeissmanZ, McBrideHJ, StillmanDJ, et al. (2000) Degradation of the transcription factor Gcn4 requires the kinase Pho85 and the SCFCDC4 ubiquitin-ligase complex. Mol Biol Cell 11 : 915–927.

13. ZhangF, GaurNA, HasekJ, KimSJ, QiuH, et al. (2008) Disrupting vesicular trafficking at the endosome attenuates transcriptional activation by Gcn4. Mol Cell Biol 28 : 6796–6818.

14. ChiY, HuddlestonMJ, ZhangX, YoungRA, AnnanRS, et al. (2001) Negative regulation of Gcn4 and Msn2 transcription factors by Srb10 cyclin-dependent kinase. Genes Dev 15 : 1078–1092.

15. ShemerR, MeimounA, HoltzmanT, KornitzerD (2002) Regulation of the transcription factor Gcn4 by Pho85 cyclin PCL5. Mol Cell Biol 22 : 5395–5404.

16. AviramS, SimonE, GildorT, GlaserF, KornitzerD (2008) Autophosphorylation-induced degradation of the Pho85 cyclin Pcl5 is essential for response to amino acid limitation. Mol Cell Biol 28 : 6858–6869.

17. RosoninaE, DuncanSM, ManleyJL (2012) Sumoylation of transcription factor Gcn4 facilitates its Srb10-mediated clearance from promoters in yeast. Genes Dev 26 : 350–355.

18. GillG (2005) Something about SUMO inhibits transcription. Curr Opin Genet Dev 15 : 536–541.

19. LipfordJR, SmithGT, ChiY, DeshaiesRJ (2005) A putative stimulatory role for activator turnover in gene expression. Nature 438 : 113–116.

20. GuglielmiB, van BerkumNL, KlapholzB, BijmaT, BoubeM, et al. (2004) A high resolution protein interaction map of the yeast Mediator complex. Nucleic Acids Res 32 : 5379–5391.

21. LiaoS-M, ZhangJ, JefferyDA, KoleskeAJ, ThompsonCM, et al. (1995) A kinase-cyclin pair in the RNA in the polymerase II holoenzyme. Nature 374 : 193–196.

22. GaliliG (1995) Regulation of Lysine and Threonine Synthesis. Plant Cell 7 : 899–906.

23. Jones EW, Fink GR (1982) Regulation of amino acid and nucleotide biosynthesis in yeast. In: "The molecular biology of the yeast Saccharomyces Metabolism and gene expression" Strathern J N Jones E W and Broach J R eds. Cold Spring Harbor Laboratory Cold Spring Harbor NY: 181–300.

24. KingsburyJM, McCuskerJH (2010) Fungal homoserine kinase (thr1Delta) mutants are attenuated in virulence and die rapidly upon threonine starvation and serum incubation. Eukaryot Cell 9 : 729–737.

25. MountainHA, BystromAS, Tang LarsenJT, KorchC (1991) Four major transcriptional responses in the methionine/threonine biosynthetic pathway of }U}Saccharomyces cerevisiae}u}. Yeast 7 : 781–803.

26. RamosC, DelgadoMA, CalderonIL (1991) Inhibition by different amino acids of the aspartate kinase and the homoserine kinase of the yeast Saccharomyces cerevisiae. FEBS Lett 278 : 123–126.

27. Arevalo-RodriguezM, PanX, BoekeJD, HeitmanJ (2004) FKBP12 controls aspartate pathway flux in Saccharomyces cerevisiae to prevent toxic intermediate accumulation. Eukaryot Cell 3 : 1287–1296.

28. AlarconCM, HeitmanJ (1997) FKBP12 physically and functionally interacts with aspartokinase in Saccharomyces cerevisiae. Mol Cell Biol 17 : 5968–5975.

29. KingsburyJM, McCuskerJH (2010) Homoserine toxicity in Saccharomyces cerevisiae and Candida albicans homoserine kinase (thr1Delta) mutants. Eukaryot Cell 9 : 717–728.

30. KimSJ, SwansonMJ, QiuH, GovindCK, HinnebuschAG (2005) Activator Gcn4p and Cyc8p/Tup1p are interdependent for promoter occupancy at ARG1 in vivo. Mol Cell Biol 25 : 11171–11183.

31. HinnebuschAG (1988) Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae. Microbiol Rev 52 : 248–273.

32. DeLaBarreB, ThompsonPR, WrightGD, BerghuisAM (2000) Crystal structures of homoserine dehydrogenase suggest a novel catalytic mechanism for oxidoreductases. Nat Struct Biol 7 : 238–244.

33. HinnebuschAG (1985) A hierarchy of trans-acting factors modulate translation of an activator of amino acid biosynthetic genes in Saccharomyces cerevisiae. Mol Cell Biol 5 : 2349–2360.

34. MuellerPP, HinnebuschAG (1986) Multiple upstream AUG codons mediate translational control of GCN4. Cell 45 : 201–207.

35. TanseyWP (2001) Transcriptional activation: risky business. Genes Dev 15 : 1045–1050.

36. DrysdaleCM, DueñasE, JacksonBM, ReusserU, BrausGH, et al. (1995) The transcriptional activator GCN4 contains multiple activation domains that are critically dependent on hydrophobic amino acids. Mol Cell Biol 15 : 1220–1233.

37. Hinnebusch AG (1992) General and pathway-specific regulatory mechanisms controlling the synthesis of amino acid biosynthetic enzymes in Saccharomyces cerevisiae. In: Broach JR, Jones EW, Pringle JR, editors. The Molecular and Cellular Biology of the Yeast Saccharomyces: Gene Expression. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. pp. 319–414.

38. ZhangF, SumibcayL, HinnebuschAG, SwansonMJ (2004) A triad of subunits from the Gal11/tail domain of Srb mediator is an in vivo target of transcriptional activator Gcn4p. Mol Cell Biol 24 : 6871–6886.

39. YamakiH, YamaguchiM, TsuruoT, YamaguchiH (1992) Mechanism of action of an antifungal antibiotic, RI-331, (S) 2-amino-4-oxo-5-hydroxypentanoic acid; kinetics of inactivation of homoserine dehydrogenase from Saccharomyces cerevisiae. J Antibiot (Tokyo) 45 : 750–755.

40. GoldsteinAL, McCuskerJH (1999) Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae. Yeast 15 : 1541–1553.

41. MoehleCM, HinnebuschAG (1991) Association of RAP1 binding sites with stringent control of ribosomal protein gene transcription in Saccharomyces cerevisiae. Mol Cell Biol 11 : 2723–2735.

42. SchmittME, BrownTA, TrumpowerBL (1990) A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res 18 : 3091–3092.

43. GaurNA, HasekJ, BricknerDG, QiuH, ZhangF, et al. (2013) Vps factors are required for efficient transcription elongation in budding yeast. Genetics 193 : 829–851.

44. ReidGA, SchatzG (1982) Import of proteins into mitochondria. Yeast cells grown in the presence of carbonyl cyanide m-chlorophenylhydrazone accumulate massive amounts of some mitochondrial precursor polypeptides. J Biol Chem 257 : 13056–13061.

45. CiganAM, FoianiM, HannigEM, HinnebuschAG (1991) Complex formation by positive and negative translational regulators of GCN4. Mol Cell Biol 11 : 3217–3228.

46. KornitzerD (2002) Monitoring protein degradation. Methods Enzymol 351 : 639–647.

47. MontpetitB, ThomsenND, HelmkeKJ, SeeligerMA, BergerJM, et al. (2011) A conserved mechanism of DEAD-box ATPase activation by nucleoporins and InsP6 in mRNA export. Nature 472 : 238–242.

48. SattleggerE, SwansonMJ, AshcraftEA, JenningsJL, FeketeRA, et al. (2004) YIH1 is an actin-binding protein that inhibits protein kinase GCN2 and impairs general amino acid control when overexpressed. J Biol Chem 279 : 29952–29962.

49. GietzRD, SuginoA (1988) New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74 : 527–534.

50. SikorskiRS, HieterP (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122 : 19–27.

51. HinnebuschAG, LucchiniG, FinkGR (1985) A synthetic HIS4 regulatory element confers general amino acid control on the cytochrome c gene CYC1 of yeast. Proc Natl Acad Sci U S A 82 : 498–502.

52. DonahueTF, CiganAM (1988) Genetic selection for mutations that reduce or abolish ribosomal recognition of the HIS4 translational initiator region. Mol Cell Biol 8 : 2955–2963.

53. LongtineMS, McKenzie IIIA, DemariniDJ, ShahNG, WachA, et al. (1998) Additonal modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14 : 953–961.

54. YoonS, QiuH, SwansonMJ, HinnebuschAG (2003) Recruitment of SWI/SNF by Gcn4p does not require Snf2p or Gcn5p but depends strongly on SWI/SNF integrity, SRB mediator, and SAGA. Mol Cell Biol 23 : 8829–8845.