Amino Acids

, Volume 40, Issue 2, pp 623–631 | Cite as

Mcm1p binding sites in the ARG1 promoter positively regulate ARG1 transcription and S. cerevisiae growth in the absence of arginine and Gcn4p

Original Article

Abstract

In this study, we investigated the activating role of Mcm1p at ARG1 during arginine starvation. Our results showed that two Mcm1p binding sites positively contribute to ARG1 transcription and cell growth. Especially, we provide strong evidence that the Mcm1p binding sites play a positive role in ARG1 transcription to overcome arginine starvation in the absence of Gcn4p. In addition, we found that the Mcm1p binding sites are not only regulated by the presence or absence of arginine but also in the presence or absence of other amino acids. These findings suggest that the ARG1 promoter utilizes different DNA elements to control transcription, depending on which amino acids are detected in the medium.

Keywords

Gcn4p Mcm1p ARG1 SD and SC media Arginine starvation 

Abbreviations

SD

Synthetic defined minimal

SC

Synthetic defined complete

ARC

Arginine control

MADS

MCM1, AGAMOUS, DEFICIENS, and serum response factor

WT

Wild type

ARG

Arginine

URA

Uracil

Can

Canavanine

Notes

Acknowledgments

We thank Marjolaine Crabeel for the SS5 strain and M13mp7-ARG1. This work was supported by Basic Science Program through the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2009-0070791).

Supplementary material

726_2010_687_MOESM1_ESM.tif (942 kb)
Supplementary Figure 1. Two Mcm1p binding sites have suppressive function in GCN4 deletion strain for survival during arginine starvation. (A and B) The gcn4ΔSS5 strains containing mutated ARG1 alleles are indicated on the left. (See Table 1 for description of strains.) “No” indicates WT ARG1 promoter in gcn4Δ SS5 strains. Cells were grown to saturation in SD+URA medium at 30°C, serially diluted tenfold, and spotted onto SD+ARG, SD, SD+Can (Canavanine; 0.05 μg/ml), or SC-ARG supplemented with uracil. The growth phenotype following more than two days’incubation at 30°C is shown. (TIFF 941 kb)
726_2010_687_MOESM2_ESM.doc (42 kb)
Supplementary material 2 (DOC 41 kb)

References

  1. Carr EA, Mead J, Vershon AK (2004) Alpha1-induced DNA bending is required for transcriptional activation by the Mcm1-alpha1 complex. Nucleic Acids Res 32:2298–2305CrossRefPubMedGoogle Scholar
  2. Crabeel M, de Rijcke M, Seneca S, Heimberg H, Pfeiffer I, Matisova A (1995) Further definition of the sequence and position requirements of the arginine control element that mediates repression and induction by arginine in Saccharomyces cerevisiae. Yeast 11:1367–1380CrossRefPubMedGoogle Scholar
  3. Darieva Z, Pic-Taylor A, Boros J, Spanos A, Geymonat M, Reece RJ, Sedgwick SG, Sharrocks AD, Morgan BA (2003) Cell cycle-regulated transcription through the FHA domain of Fkh2p and the coactivator Ndd1p. Curr Biol 13:1740–1745CrossRefPubMedGoogle Scholar
  4. De Rijcke M, Seneca S, Punyammalee B, Glansdorff N, Crabeel M (1992) Characterization of the DNA target site for the yeast ARGR regulatory complex, a sequence able to mediate repression or induction by arginine. Mol Cell Biol 12:68–81PubMedGoogle Scholar
  5. Gavin IM, Kladde MP, Simpson RT (2000) Tup1p represses Mcm1p transcriptional activation and chromatin remodeling of an a-cell-specific gene. EMBO J 19:5875–5883CrossRefPubMedGoogle Scholar
  6. Hinnebusch AG (1992) The molecular and cellular biology of the yeast saccharomyces: gene expression. In: Broach JR, Jones EW, Pringle JR (eds) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 319–414Google Scholar
  7. Messenguy F, Dubois E (1993) Genetic evidence for a role for MCM1 in the regulation of arginine metabolism in Saccharomyces cerevisiae. Mol Cell Biol 13:2586–2592PubMedGoogle Scholar
  8. Messenguy F, Dubois E (2003) Role of MADS box proteins and their cofactors in combinatorial control of gene expression and cell development. Gene 316:1–21CrossRefPubMedGoogle Scholar
  9. Natarajan K, Meyer MR, Jackson BM, Slade D, Roberts C, Hinnebusch AG, Marton MJ (2001) Transcriptional profiling shows that Gcn4p is a master regulator of gene expression during amino acid starvation in yeast. Mol Cell Biol 21:4347–4368CrossRefPubMedGoogle Scholar
  10. Penn MD, Galgoci B, Greer H (1983) Identification of AAS genes and their regulatory role in general control of amino acids biosynthesis in yeast. Proc Natl Acad Sci USA 80:2704–2708CrossRefPubMedGoogle Scholar
  11. Qiu H, Hu C, Yoon S, Natarajan K, Swanson MJ, Hinnebusch AG (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 24:4104–4117CrossRefPubMedGoogle Scholar
  12. Sherman F (1991) Getting started with yeast. Methods Enzymol 194:3–21CrossRefPubMedGoogle Scholar
  13. Tan S, Richmond TJ (1998) Crystal structure of the yeast MATalpha2/MCM1/DNA ternary complex. Nature 391:660–666CrossRefPubMedGoogle Scholar
  14. Yoon S, Hinnebusch AG (2009) Mcm1p binding sites in ARG1 positively regulate Gcn4p binding and SWI/SNF recruitment. Biochem Biophys Res Commun 381(1):123–128CrossRefPubMedGoogle Scholar
  15. Yoon S, Qiu H, Swanson MJ, Hinnebusch AG (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 23:8829–8845CrossRefPubMedGoogle Scholar
  16. Yoon S, Govind CK, Qiu H, Kim SJ, Dong J, Hinnebusch AG (2004) Recruitment of the ArgR/Mcm1p repressor is stimulated by the activator Gcn4p: a self-checking activation mechanism. Proc Natl Acad Sci USA 91:11713–11718CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Research InstituteNational Cancer CenterGoyang-siRepublic of Korea
  2. 2.Lee Gil Ya Cancer and Diabetes InstituteGachon University of Medicine and ScienceIncheonRepublic of Korea

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