Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 2001 Mar;157(3):1205–1215. doi: 10.1093/genetics/157.3.1205

Transcriptional regulators of the Schizosaccharomyces pombe fbp1 gene include two redundant Tup1p-like corepressors and the CCAAT binding factor activation complex.

R T Janoo 1, L A Neely 1, B R Braun 1, S K Whitehall 1, C S Hoffman 1
PMCID: PMC1461578  PMID: 11238405

Abstract

The Schizosaccharomyces pombe fbp1 gene, which encodes fructose-1,6-bis-phosphatase, is transcriptionally repressed by glucose through the activation of the cAMP-dependent protein kinase A (PKA) and transcriptionally activated by glucose starvation through the activation of a mitogen-activated protein kinase (MAPK). To identify transcriptional regulators acting downstream from or in parallel to PKA, we screened an adh-driven cDNA plasmid library for genes that increase fbp1 transcription in a strain with elevated PKA activity. Two such clones express amino-terminally truncated forms of the S. pombe tup12 protein that resembles the Saccharomyces cerevisiae Tup1p global corepressor. These clones appear to act as dominant negative alleles. Deletion of both tup12 and the closely related tup11 gene causes a 100-fold increase in fbp1-lacZ expression, indicating that tup11 and tup12 are redundant negative regulators of fbp1 transcription. In strains lacking tup11 and tup12, the atf1-pcr1 transcriptional activator continues to play a central role in fbp1-lacZ expression; however, spc1 MAPK phosphorylation of atf1 is no longer essential for its activation. We discuss possible models for the role of tup11- and tup12-mediated repression with respect to signaling from the MAPK and PKA pathways. A third clone identified in our screen expresses the php5 protein subunit of the CCAAT-binding factor (CBF). Deletion of php5 reduces fbp1 expression under both repressed and derepressed conditions. The CBF appears to act in parallel to atf1-pcr1, although it is unclear whether or not CBF activity is regulated by PKA.

Full Text

The Full Text of this article is available as a PDF (538.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Byrne S. M., Hoffman C. S. Six git genes encode a glucose-induced adenylate cyclase activation pathway in the fission yeast Schizosaccharomyces pombe. J Cell Sci. 1993 Aug;105(Pt 4):1095–1100. doi: 10.1242/jcs.105.4.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bähler J., Wu J. Q., Longtine M. S., Shah N. G., McKenzie A., 3rd, Steever A. B., Wach A., Philippsen P., Pringle J. R. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast. 1998 Jul;14(10):943–951. doi: 10.1002/(SICI)1097-0061(199807)14:10<943::AID-YEA292>3.0.CO;2-Y. [DOI] [PubMed] [Google Scholar]
  3. Caspari T. Onset of gluconate-H+ symport in Schizosaccharomyces pombe is regulated by the kinases Wis1 and Pka1, and requires the gti1+ gene product. J Cell Sci. 1997 Oct;110(Pt 20):2599–2608. doi: 10.1242/jcs.110.20.2599. [DOI] [PubMed] [Google Scholar]
  4. Dal Santo P., Blanchard B., Hoffman C. S. The Schizosaccharomyces pombe pyp1 protein tyrosine phosphatase negatively regulates nutrient monitoring pathways. J Cell Sci. 1996 Jul;109(Pt 7):1919–1925. doi: 10.1242/jcs.109.7.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Degols G., Russell P. Discrete roles of the Spc1 kinase and the Atf1 transcription factor in the UV response of Schizosaccharomyces pombe. Mol Cell Biol. 1997 Jun;17(6):3356–3363. doi: 10.1128/mcb.17.6.3356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Degols G., Shiozaki K., Russell P. Activation and regulation of the Spc1 stress-activated protein kinase in Schizosaccharomyces pombe. Mol Cell Biol. 1996 Jun;16(6):2870–2877. doi: 10.1128/mcb.16.6.2870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ducker C. E., Simpson R. T. The organized chromatin domain of the repressed yeast a cell-specific gene STE6 contains two molecules of the corepressor Tup1p per nucleosome. EMBO J. 2000 Feb 1;19(3):400–409. doi: 10.1093/emboj/19.3.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edmondson D. G., Smith M. M., Roth S. Y. Repression domain of the yeast global repressor Tup1 interacts directly with histones H3 and H4. Genes Dev. 1996 May 15;10(10):1247–1259. doi: 10.1101/gad.10.10.1247. [DOI] [PubMed] [Google Scholar]
  9. Edmondson D. G., Zhang W., Watson A., Xu W., Bone J. R., Yu Y., Stillman D., Roth S. Y. In vivo functions of histone acetylation/deacetylation in Tup1p repression and Gcn5p activation. Cold Spring Harb Symp Quant Biol. 1998;63:459–468. doi: 10.1101/sqb.1998.63.459. [DOI] [PubMed] [Google Scholar]
  10. Elledge S. J., Zhou Z., Allen J. B., Navas T. A. DNA damage and cell cycle regulation of ribonucleotide reductase. Bioessays. 1993 May;15(5):333–339. doi: 10.1002/bies.950150507. [DOI] [PubMed] [Google Scholar]
  11. Estruch F., Carlson M. Two homologous zinc finger genes identified by multicopy suppression in a SNF1 protein kinase mutant of Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jul;13(7):3872–3881. doi: 10.1128/mcb.13.7.3872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Forsburg S. L., Guarente L. Identification and characterization of HAP4: a third component of the CCAAT-bound HAP2/HAP3 heteromer. Genes Dev. 1989 Aug;3(8):1166–1178. doi: 10.1101/gad.3.8.1166. [DOI] [PubMed] [Google Scholar]
  13. Hai T. W., Liu F., Allegretto E. A., Karin M., Green M. R. A family of immunologically related transcription factors that includes multiple forms of ATF and AP-1. Genes Dev. 1988 Oct;2(10):1216–1226. doi: 10.1101/gad.2.10.1216. [DOI] [PubMed] [Google Scholar]
  14. Hoffman C. S., Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. doi: 10.1016/0378-1119(87)90131-4. [DOI] [PubMed] [Google Scholar]
  15. Hoffman C. S., Winston F. A transcriptionally regulated expression vector for the fission yeast Schizosaccharomyces pombe. Gene. 1989 Dec 14;84(2):473–479. doi: 10.1016/0378-1119(89)90523-4. [DOI] [PubMed] [Google Scholar]
  16. Hoffman C. S., Winston F. Glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene occurs by a cAMP signaling pathway. Genes Dev. 1991 Apr;5(4):561–571. doi: 10.1101/gad.5.4.561. [DOI] [PubMed] [Google Scholar]
  17. Isshiki T., Mochizuki N., Maeda T., Yamamoto M. Characterization of a fission yeast gene, gpa2, that encodes a G alpha subunit involved in the monitoring of nutrition. Genes Dev. 1992 Dec;6(12B):2455–2462. doi: 10.1101/gad.6.12b.2455. [DOI] [PubMed] [Google Scholar]
  18. Jin M., Fujita M., Culley B. M., Apolinario E., Yamamoto M., Maundrell K., Hoffman C. S. sck1, a high copy number suppressor of defects in the cAMP-dependent protein kinase pathway in fission yeast, encodes a protein homologous to the Saccharomyces cerevisiae SCH9 kinase. Genetics. 1995 Jun;140(2):457–467. doi: 10.1093/genetics/140.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kanoh J., Watanabe Y., Ohsugi M., Iino Y., Yamamoto M. Schizosaccharomyces pombe gad7+ encodes a phosphoprotein with a bZIP domain, which is required for proper G1 arrest and gene expression under nitrogen starvation. Genes Cells. 1996 Apr;1(4):391–408. doi: 10.1046/j.1365-2443.1996.d01-247.x. [DOI] [PubMed] [Google Scholar]
  20. Keleher C. A., Redd M. J., Schultz J., Carlson M., Johnson A. D. Ssn6-Tup1 is a general repressor of transcription in yeast. Cell. 1992 Feb 21;68(4):709–719. doi: 10.1016/0092-8674(92)90146-4. [DOI] [PubMed] [Google Scholar]
  21. Kon N., Schroeder S. C., Krawchuk M. D., Wahls W. P. Regulation of the Mts1-Mts2-dependent ade6-M26 meiotic recombination hot spot and developmental decisions by the Spc1 mitogen-activated protein kinase of fission yeast. Mol Cell Biol. 1998 Dec;18(12):7575–7583. doi: 10.1128/mcb.18.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Landry S., Pettit M. T., Apolinario E., Hoffman C. S. The fission yeast git5 gene encodes a Gbeta subunit required for glucose-triggered adenylate cyclase activation. Genetics. 2000 Apr;154(4):1463–1471. doi: 10.1093/genetics/154.4.1463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lipke P. N., Hull-Pillsbury C. Flocculation of Saccharomyces cerevisiae tup1 mutants. J Bacteriol. 1984 Aug;159(2):797–799. doi: 10.1128/jb.159.2.797-799.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lundin M., Nehlin J. O., Ronne H. Importance of a flanking AT-rich region in target site recognition by the GC box-binding zinc finger protein MIG1. Mol Cell Biol. 1994 Mar;14(3):1979–1985. doi: 10.1128/mcb.14.3.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lutfiyya L. L., Iyer V. R., DeRisi J., DeVit M. J., Brown P. O., Johnston M. Characterization of three related glucose repressors and genes they regulate in Saccharomyces cerevisiae. Genetics. 1998 Dec;150(4):1377–1391. doi: 10.1093/genetics/150.4.1377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lutfiyya L. L., Johnston M. Two zinc-finger-containing repressors are responsible for glucose repression of SUC2 expression. Mol Cell Biol. 1996 Sep;16(9):4790–4797. doi: 10.1128/mcb.16.9.4790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Maeda T., Mochizuki N., Yamamoto M. Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7814–7818. doi: 10.1073/pnas.87.20.7814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maeda T., Watanabe Y., Kunitomo H., Yamamoto M. Cloning of the pka1 gene encoding the catalytic subunit of the cAMP-dependent protein kinase in Schizosaccharomyces pombe. J Biol Chem. 1994 Apr 1;269(13):9632–9637. [PubMed] [Google Scholar]
  29. Mannervik M., Nibu Y., Zhang H., Levine M. Transcriptional coregulators in development. Science. 1999 Apr 23;284(5414):606–609. doi: 10.1126/science.284.5414.606. [DOI] [PubMed] [Google Scholar]
  30. Martínez-Pastor M. T., Marchler G., Schüller C., Marchler-Bauer A., Ruis H., Estruch F. The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). EMBO J. 1996 May 1;15(9):2227–2235. [PMC free article] [PubMed] [Google Scholar]
  31. McNabb D. S., Xing Y., Guarente L. Cloning of yeast HAP5: a novel subunit of a heterotrimeric complex required for CCAAT binding. Genes Dev. 1995 Jan 1;9(1):47–58. doi: 10.1101/gad.9.1.47. [DOI] [PubMed] [Google Scholar]
  32. Millar J. B., Buck V., Wilkinson M. G. Pyp1 and Pyp2 PTPases dephosphorylate an osmosensing MAP kinase controlling cell size at division in fission yeast. Genes Dev. 1995 Sep 1;9(17):2117–2130. doi: 10.1101/gad.9.17.2117. [DOI] [PubMed] [Google Scholar]
  33. Mukai Y., Harashima S., Oshima Y. AAR1/TUP1 protein, with a structure similar to that of the beta subunit of G proteins, is required for a1-alpha 2 and alpha 2 repression in cell type control of Saccharomyces cerevisiae. Mol Cell Biol. 1991 Jul;11(7):3773–3779. doi: 10.1128/mcb.11.7.3773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mukai Y., Matsuo E., Roth S. Y., Harashima S. Conservation of histone binding and transcriptional repressor functions in a Schizosaccharomyces pombe Tup1p homolog. Mol Cell Biol. 1999 Dec;19(12):8461–8468. doi: 10.1128/mcb.19.12.8461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Márquez J. A., Pascual-Ahuir A., Proft M., Serrano R. The Ssn6-Tup1 repressor complex of Saccharomyces cerevisiae is involved in the osmotic induction of HOG-dependent and -independent genes. EMBO J. 1998 May 1;17(9):2543–2553. doi: 10.1093/emboj/17.9.2543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Neely L. A., Hoffman C. S. Protein kinase A and mitogen-activated protein kinase pathways antagonistically regulate fission yeast fbp1 transcription by employing different modes of action at two upstream activation sites. Mol Cell Biol. 2000 Sep;20(17):6426–6434. doi: 10.1128/mcb.20.17.6426-6434.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nehlin J. O., Ronne H. Yeast MIG1 repressor is related to the mammalian early growth response and Wilms' tumour finger proteins. EMBO J. 1990 Sep;9(9):2891–2898. doi: 10.1002/j.1460-2075.1990.tb07479.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nocero M., Isshiki T., Yamamoto M., Hoffman C. S. Glucose repression of fbp1 transcription of Schizosaccharomyces pombe is partially regulated by adenylate cyclase activation by a G protein alpha subunit encoded by gpa2 (git8). Genetics. 1994 Sep;138(1):39–45. doi: 10.1093/genetics/138.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Olesen J., Hahn S., Guarente L. Yeast HAP2 and HAP3 activators both bind to the CYC1 upstream activation site, UAS2, in an interdependent manner. Cell. 1987 Dec 24;51(6):953–961. doi: 10.1016/0092-8674(87)90582-4. [DOI] [PubMed] [Google Scholar]
  40. Papamichos-Chronakis M., Conlan R. S., Gounalaki N., Copf T., Tzamarias D. Hrs1/Med3 is a Cyc8-Tup1 corepressor target in the RNA polymerase II holoenzyme. J Biol Chem. 2000 Mar 24;275(12):8397–8403. doi: 10.1074/jbc.275.12.8397. [DOI] [PubMed] [Google Scholar]
  41. Proft M., Serrano R. Repressors and upstream repressing sequences of the stress-regulated ENA1 gene in Saccharomyces cerevisiae: bZIP protein Sko1p confers HOG-dependent osmotic regulation. Mol Cell Biol. 1999 Jan;19(1):537–546. doi: 10.1128/mcb.19.1.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ptashne M., Gann A. Transcriptional activation by recruitment. Nature. 1997 Apr 10;386(6625):569–577. doi: 10.1038/386569a0. [DOI] [PubMed] [Google Scholar]
  43. Roesler W. J., Vandenbark G. R., Hanson R. W. Cyclic AMP and the induction of eukaryotic gene transcription. J Biol Chem. 1988 Jul 5;263(19):9063–9066. [PubMed] [Google Scholar]
  44. Samejima I., Mackie S., Fantes P. A. Multiple modes of activation of the stress-responsive MAP kinase pathway in fission yeast. EMBO J. 1997 Oct 15;16(20):6162–6170. doi: 10.1093/emboj/16.20.6162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Samejima I., Mackie S., Warbrick E., Weisman R., Fantes P. A. The fission yeast mitotic regulator win1+ encodes an MAP kinase kinase kinase that phosphorylates and activates Wis1 MAP kinase kinase in response to high osmolarity. Mol Biol Cell. 1998 Aug;9(8):2325–2335. doi: 10.1091/mbc.9.8.2325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Schmitt A. P., McEntee K. Msn2p, a zinc finger DNA-binding protein, is the transcriptional activator of the multistress response in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5777–5782. doi: 10.1073/pnas.93.12.5777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Schultz J., Carlson M. Molecular analysis of SSN6, a gene functionally related to the SNF1 protein kinase of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Oct;7(10):3637–3645. doi: 10.1128/mcb.7.10.3637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Schultz J., Marshall-Carlson L., Carlson M. The N-terminal TPR region is the functional domain of SSN6, a nuclear phosphoprotein of Saccharomyces cerevisiae. Mol Cell Biol. 1990 Sep;10(9):4744–4756. doi: 10.1128/mcb.10.9.4744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Shieh J. C., Wilkinson M. G., Buck V., Morgan B. A., Makino K., Millar J. B. The Mcs4 response regulator coordinately controls the stress-activated Wak1-Wis1-Sty1 MAP kinase pathway and fission yeast cell cycle. Genes Dev. 1997 Apr 15;11(8):1008–1022. doi: 10.1101/gad.11.8.1008. [DOI] [PubMed] [Google Scholar]
  50. Shiozaki K., Russell P. Conjugation, meiosis, and the osmotic stress response are regulated by Spc1 kinase through Atf1 transcription factor in fission yeast. Genes Dev. 1996 Sep 15;10(18):2276–2288. doi: 10.1101/gad.10.18.2276. [DOI] [PubMed] [Google Scholar]
  51. Shiozaki K., Shiozaki M., Russell P. Mcs4 mitotic catastrophe suppressor regulates the fission yeast cell cycle through the Wik1-Wis1-Spc1 kinase cascade. Mol Biol Cell. 1997 Mar;8(3):409–419. doi: 10.1091/mbc.8.3.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Smith R. L., Johnson A. D. A sequence resembling a peroxisomal targeting sequence directs the interaction between the tetratricopeptide repeats of Ssn6 and the homeodomain of alpha 2. Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3901–3906. doi: 10.1073/pnas.070506797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Stettler S., Warbrick E., Prochnik S., Mackie S., Fantes P. The wis1 signal transduction pathway is required for expression of cAMP-repressed genes in fission yeast. J Cell Sci. 1996 Jul;109(Pt 7):1927–1935. doi: 10.1242/jcs.109.7.1927. [DOI] [PubMed] [Google Scholar]
  54. Takeda T., Toda T., Kominami K., Kohnosu A., Yanagida M., Jones N. Schizosaccharomyces pombe atf1+ encodes a transcription factor required for sexual development and entry into stationary phase. EMBO J. 1995 Dec 15;14(24):6193–6208. doi: 10.1002/j.1460-2075.1995.tb00310.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tanaka N., Ohuchi N., Mukai Y., Osaka Y., Ohtani Y., Tabuchi M., Bhuiyan M. S., Fukui H., Harashima S., Takegawa K. Isolation and characterization of an invertase and its repressor genes from Schizosaccharomyces pombe. Biochem Biophys Res Commun. 1998 Apr 7;245(1):246–253. doi: 10.1006/bbrc.1998.8406. [DOI] [PubMed] [Google Scholar]
  56. Tzamarias D., Struhl K. Functional dissection of the yeast Cyc8-Tup1 transcriptional co-repressor complex. Nature. 1994 Jun 30;369(6483):758–761. doi: 10.1038/369758a0. [DOI] [PubMed] [Google Scholar]
  57. Wahi M., Komachi K., Johnson A. D. Gene regulation by the yeast Ssn6-Tup1 corepressor. Cold Spring Harb Symp Quant Biol. 1998;63:447–457. doi: 10.1101/sqb.1998.63.447. [DOI] [PubMed] [Google Scholar]
  58. Warbrick E., Fantes P. A. The wis1 protein kinase is a dosage-dependent regulator of mitosis in Schizosaccharomyces pombe. EMBO J. 1991 Dec;10(13):4291–4299. doi: 10.1002/j.1460-2075.1991.tb05007.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Watanabe Y., Lino Y., Furuhata K., Shimoda C., Yamamoto M. The S.pombe mei2 gene encoding a crucial molecule for commitment to meiosis is under the regulation of cAMP. EMBO J. 1988 Mar;7(3):761–767. doi: 10.1002/j.1460-2075.1988.tb02873.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Welton R. M., Hoffman C. S. Glucose monitoring in fission yeast via the Gpa2 galpha, the git5 Gbeta and the git3 putative glucose receptor. Genetics. 2000 Oct;156(2):513–521. doi: 10.1093/genetics/156.2.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wilkinson M. G., Samuels M., Takeda T., Toone W. M., Shieh J. C., Toda T., Millar J. B., Jones N. The Atf1 transcription factor is a target for the Sty1 stress-activated MAP kinase pathway in fission yeast. Genes Dev. 1996 Sep 15;10(18):2289–2301. doi: 10.1101/gad.10.18.2289. [DOI] [PubMed] [Google Scholar]
  62. Williams F. E., Varanasi U., Trumbly R. J. The CYC8 and TUP1 proteins involved in glucose repression in Saccharomyces cerevisiae are associated in a protein complex. Mol Cell Biol. 1991 Jun;11(6):3307–3316. doi: 10.1128/mcb.11.6.3307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Wu J., Trumbly R. J. Multiple regulatory proteins mediate repression and activation by interaction with the yeast Mig1 binding site. Yeast. 1998 Aug;14(11):985–1000. doi: 10.1002/(SICI)1097-0061(199808)14:11<985::AID-YEA294>3.0.CO;2-C. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES