Molecular and General Genetics MGG

, Volume 243, Issue 3, pp 315–324 | Cite as

Pleiotropic function of ArgRIIIp (Arg82p), one of the regulators of arginine metabolism in Saccharomyces cerevisiae. Role in expression of cell-type-specific genes

  • E. Dubois
  • F. Messenguy
Original Paper


ArgRIIIp (Arg82p), together with ArgRIp (Arg80p), ArgRIIp (Arg81p) and Mcmlp, regulates the expression of arginine anabolic and catabolic genes. An argRIII mutant constitutively expresses five anabolic enzymes and is impaired in the induction of the synthesis of two catabolic enzymes. A genomic disruption of the ARGRIII gene not only leads to an argR phenotype, but also prevents cell growth at 37°C. The disrupted strain is sterile especially in an α background and transcription of α- and a-specific genes (MFα1 and STE2) is strongly reduced. By gel retardation assays we show that the binding of the Mcmlp present in a crude protein extract from an argRIII mutant strain to the P(PAL) sequence is impaired. Sporulation of α/aargRIII:: URA3 homozygous diploids is also affected. Overexpression of Mcm1p in an argRIII-disrupted strain restores the mating competence of the strain, the ability to form a protein complex with P(PAL) DNA in vitro, and the regulation of arginine metabolism. However, overexpression of Mcm1p does not complement the sporulation deficiency of the argRIII-disrupted strain, nor does it complement its growth defect at 37°C. Western blot analysis indicates that Mcm1p is less abundant in a strain devoid of ArgRIIIp than in wild type.

Key words

Yeast Arginine Cell-type regulation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alwine JC, Kemp DJ, Stark GR (1977) Detection of specific fragments of DNA by fractionation of gels and transfer to diazobenzylmethyl paper. Proc Natl Acad Sci USA 74:5350–5354Google Scholar
  2. Ammerer G (1990) Identification, purification and cloning of a polypeptide (PRTF/GRM) that binds to mating specific promoter elements in yeast. Genes Dev 4:299–312Google Scholar
  3. Béchet J, Grenson M, Wiame JM (1970) Mutations affecting the repressibility of arginine biosynthetic enzymes in S. cerevisiae. Eur J Biochem 12:31–39Google Scholar
  4. Bender A, Sprague GF (1987) MATα1 protein, a yeast transcription activator binds synergistically with a second protein to a set of cell-type-specific genes. Cell 50:681–690Google Scholar
  5. Bercy J, Dubois E, Messenguy F (1987) Regulation of arginine metabolism in S. cerevisiae: expression of the three ARGR regulatory genes and cellular localization of their products. Gene 55:277–285Google Scholar
  6. Bonneaud N, Ozier-Kalogeropoulos O, Li G, Labouesse M, Minvielle-Sebastia L, Lacroute F (1991) A family of low and high copy replicative, integrative and single-stranded S. cerevisiae/E. coli shuttle vectors. Yeast 7:609–615Google Scholar
  7. Christ C (1992) Characterization of MCM1, a multifunctional yeast DNA binding protein. PhD Thesis, Cornell UniversityGoogle Scholar
  8. Christ C, Tye BK (1991) Functional domains of the yeast transcription/replication factor MCM1. Genes Dev 5:751–763Google Scholar
  9. Delforge J, Messenguy F, Wiame JM (1975) The regulation of arginine biosynthesis in S. cerevisiae: the specificity of argR mutations and the general control of amino acid biosynthesis. Eur J Biochem 57:231–239Google Scholar
  10. Dubois E, Messenguy F (1991) In vitro studies of the binding of the ARGR protein to the ARGR5, 6 promoter. Mol Cell Biol 11:2162–2168Google Scholar
  11. Dubois E, Bercy J, Messenguy F (1987) Characterization of two genes ARGRI and ARGRIII required for specific regulation of arginine metabolism, in yeast. Mol Gen Genet 207:142–148Google Scholar
  12. Elble R, Tye BK (1991) Both activation and repression of a-matingtype-specific genes in yeast require transcription factor MCM1. Proc Natl Acad Sci USA 88:10966–10970Google Scholar
  13. Elble R, Tye BK (1992) Chromosome loss, hyperrecombination, and cell cycle arrest in a yeast mcm1 mutant. Mol Biol Cell 3:971–980Google Scholar
  14. Errede B, Ammerer G (1989) STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast is part of protein-DNA complexes. Genes Dev 3:1349–1361Google Scholar
  15. Garnier J (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120:97–120Google Scholar
  16. Herskowitz I (1988) Life cycle of the budding yeast Saccharomyces cerevisiae. Microbiol Rev 52:536–563Google Scholar
  17. Jarvis EE, Clark KL, Sprague GF (1989) The yeast transcription activator PRTF, a homolog of the mammalian seEzEσrum-response factor is encoded by the MCM1 gene. Genes Dev 3:936–945Google Scholar
  18. Keleher CA, Goutte C, Johnson AD (1988) The yeast cell-typespecific repressor α2 acts cooperatively with a non-cell-type-specific protein. Cell 53:927–936Google Scholar
  19. Keleher CA, Passemore S, Johnson AD (1989) Yeast repressor α2 binds to its operator cooperatively with yeast protein MCM1. Mol Cell Biol 9:5228–5230Google Scholar
  20. Keleher CA, Redd MJ, Schultz J, Carlson M, Johnson AD (1992) Ssn6-Tup1 is a general repressor of transcription in yeast. Cell 68:709–719Google Scholar
  21. Kurjan J, Herskowitz I (1982) Structure of a yeast pheromone gene (MFα): a putative α-factor precursor contains four tandem copies of mature α-factor. Cell 30:933–943Google Scholar
  22. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophase T4. Nature 217:680–685Google Scholar
  23. Lindquist S (1981) Regulation of protein synthesis during heat shock. Nature 293:311–314Google Scholar
  24. Malone EA, Clark CD, Chiang A, Winston F (1991) Mutations in SPT16/CDC68 suppresses cis- and trans-acting mutations that affect promoter function in S. cerevisiae. Mol Cell Biol 11:5710–5717Google Scholar
  25. Maniatis T, Fritsch EF, Sambrok J (1982) Molecular cloning: a laboratory manual. Cold Spring arbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  26. Messenguy F, Penninckx M, Wiame JM (1971) Interaction between arginase and ornithine carbamoyltransferase in S. cerevisiae. Eur J Biochem 22:277–286Google Scholar
  27. Messenguy F (1976) Regulation of arginine biosynthesis in S. cerevisiae: isolation of a cis-dominant constitutive mutant for ornithine carbamoyltransferase synthesis. J Bacteriol 128:49–55Google Scholar
  28. Messenguy F, Dubois E (1993) Participation of MCM1 in the regulation of arginine metabolism in S. cerevisiae. Mol Cell Biol 13:2586–2592Google Scholar
  29. Messenguy F, Dubois E (1983) Participation of transcriptional and post-transcriptional mechanisms in the control of arginine metabolism in yeast. Mol Gen Genet 189:148–156Google Scholar
  30. Messenguy F, Dubois E, Boonchird C (1991) Determination of the DNA binding sequences of ARGR proteins to arginine anabolic and catabolic promoters. Mol Cell Biol 11:2852–2863Google Scholar
  31. Morrison A, Miller EJ, Prakash L (1988) Domain structure and functional analysis of the carboxy-terminal polyacidic sequence of the RAD6 protein of S. cerevisiae. Mol Cell Biol 8:1179–1185Google Scholar
  32. Neal-Burnette W (1981) Western blotting: electrophoretic transfer of proteins from dodecyl sulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203Google Scholar
  33. Norman C, Runswick M, Polloc KR, Treisman R (1988) Isolation and properties of cDNA clones encoding SRF, a transcription factor that binds to the c-fos serum response element. Cell 55:989–1003Google Scholar
  34. Passemore S, Meine GT, Christ C, Tye BK (1988) S. cerevisiae protein involved in plasmid maintenance is necessary for mating of MATα cells. J Mol Biol 204:593–606Google Scholar
  35. Peterson CL, Herskowitz I (1992) Characterization of the yeast SW11, SW12, and SW13 genes, which encode a global activator of transcription. Cell 68:573–583Google Scholar
  36. Qiu HF, Dubois E, Bröen P, Messenguy F (1990) Functional analysis of ARGRI and ARGRIII regulatory proteins involved in the regulation of arginine metabolism in S. cerevisiae. Mol Gen Genet 222:192–200Google Scholar
  37. Qiu HF, Dubois E, Messenguy F (1991) Dissection of the bifunctional ARGRII protein involved in the regulation of arginine anabolic and catabolic pathways. Mol Cell Biol 11:2169–2179Google Scholar
  38. Ramos F, Wiame JM (1985) Mutation affecting the specific regulatory control of lysine biosynthetic enzymes in S. cerevisiae. Mol Gen Genet 200:291–294Google Scholar
  39. Rigby WJB, Dieckman M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specificity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251Google Scholar
  40. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  41. Smith DL, Johnson AD (1992) A molecular mechanism for combinatorial control in yeast: MCM1 protein sets the spacing and orientation of the homeodomains of an α2 dimer. Cell 68:133–142Google Scholar
  42. Sommer H, Beltran JP, Huyjser P, Pape H, Lönnig WE, Saedler H, Scharz-Sommer Z (1990) Deficiens, a homeotic gene involved in the control of flower morphogenesis in Antirrhinum majus: the protein shows homology to transcription factors. EMBO J 9:605–613Google Scholar
  43. Swanson MS, Carlson M, Winston F (1990) SPT6, an essential gene that affects transcription in S. cerevisiae, encodes a nuclear protein with an extremely acidic amino terminus. Mol Cell Biol 10:4935–4941Google Scholar
  44. Swanson MS, Malone EA, Winston F (1991) SPT5, an essential gene important for normal transcription in S. cerevisiae, encodes an acidic nuclear protein with a carboxy-terminal repeat. Mol Cell Biol 11:3009–3019Google Scholar
  45. Thuriaux P (1969) Existence de gènes régulateurs couplant la répression de la biosynthèse et l'induction du catabolisme de l'arginine dans S. cerevisiae. PhD Thesis, Brussels University, Brussels, BelgiumGoogle Scholar
  46. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 79:4350–4354Google Scholar
  47. Vidal M, Gaber RF (1991) RPD3 encodes a second factor required to achieve maximum positive and negative transcriptional states in S. cerevisiae. Mol Cell Biol 11:6317–6327Google Scholar
  48. Vidal M, Strich R, Easton Esposito R, Gaber, RF (1991) RPD1 (SINS/UME4) is required for maximal activation and repression of diverse yeast genes. Mol Cell Biol 11:6306–6316Google Scholar
  49. Yanovsky MF, Ma H, Bowman JL, Drews GN, Feldmann KA, Meyerowits EM (1990) The protein encoded by the Arabidopsis homeotic gene Agamous resembles transcription factors. Nature 346:35–39Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • E. Dubois
    • 1
    • 2
  • F. Messenguy
    • 1
    • 2
  1. 1.Institut de Recherches du CERIABrusselsBelgium
  2. 2.Laboratoire de Microbiologie de l'Université Libre de BruxellesBelgium

Personalised recommendations