Pathways and Mechanisms of Yeast Competence: A New Frontier of Yeast Genetics

  • Petar Tomev MitrikeskiEmail author
Part of the Fungal Biology book series (FUNGBIO)


Spontaneous yeast competence for exogenous DNA uptake is powered by naturally occurring cell traits/processes controlled by genetic and environmental factors. Therefore, yeast transformability is defined as a complex, quantitative genetic trait which may have contributed to fungal evolution. Such contemplation has elucidated comprehensive mechanisms of natural competence involved in spontaneous yeast transformation under environmental conditions. Moreover, several recent reports have identified many genes and/or entire cell processes responsible for the phenomenon in yeast Saccharomyces cerevisiae. However, this is hardly the case for many non-Saccharomyces species significantly important to humanity. Therefore, knowledge on pathways/mechanisms of eukaryotic competence is still only partial, inconclusive and not comprehensive. For instance, although we can line up all the necessary steps leading to transformation, we still lack fundamental knowledge to logically assemble them. Obviously, many important questions still remain open to a more systematic and targeted approach that will paradigmatically broaden our knowledge on the subject.


Yeast Saccharomyces Transformation Natural competence Pathways and Mechanisms 



I wish to thank to my postdoc boss Dr. Krunoslav Brčić-Kostić (Ruđer Bošković Institute) for critical reading of the manuscript and generous support and to M.Sc. Juraj Bergman (Ruđer Bošković Institute) for polishing the English. I would also like to acknowledge the Editors for editing the manuscript in such a way that my original ideas and thoughts are now expressed more profoundly. Finally, I specially wish to acknowledge Professor Kousaku Murata (Graduate School of Agriculture, Kyoto University) for his ideas on fungal transformation grossly influenced mine.


  1. Armaleo D, Ye GN, Klein TM, Shark KB, Sanford JC, Johnston SA (1990) Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi. Curr Genet 17:97–103PubMedCrossRefGoogle Scholar
  2. Avery OT, Macleod CM, McCarty M (1944) Studies on the chemical nature of the substance inducing transformation of pneumococcal types: induction of transformation by a desoxyribonucleic acid fraction isolated from Pneumococcus type III. J Exp Med 79:137–158PubMedCrossRefPubMedCentralGoogle Scholar
  3. Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–109PubMedCrossRefGoogle Scholar
  4. Bonnefoy N, Fox TD (2007) Directed alteration of Saccharomyces cerevisiae mitochondrial DNA by biolistic transformation and homologous recombination. Methods Mol Biol 372:153–166PubMedCrossRefPubMedCentralGoogle Scholar
  5. Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133PubMedCrossRefGoogle Scholar
  6. Bruschi CV, Comer AR, Howe GA (1987) Specificity of DNA uptake during whole cell transformation of S. cerevisiae. Yeast 3:131–137PubMedCrossRefGoogle Scholar
  7. Brzobohatý B, Kováč L (1986) Factors enhancing genetic transformation of intact yeast cells modify cell wall porosity. J Gen Microbiol 132:3089–3093PubMedGoogle Scholar
  8. Bundock P, den Dulk-Ras A, Beijersbergen A, Hooykaas PJ (1995) Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J 14:3206–3214PubMedPubMedCentralGoogle Scholar
  9. Chaustova L, Miliukienė V, Zimkus A, Razumas V (2008) Metabolic state and cell cycle as determinants of facilitated uptake of genetic information by yeast Saccharomyces cerevisiae. Cent Eur J Biol 3:417–421CrossRefGoogle Scholar
  10. Chen P, Liu HH, Cui R, Zhang ZL, Pang DW, Xie ZX, Zheng HZ, Lu ZX, Tong H (2008) Visualized investigation of yeast transformation induced with Li+ and polyethylene glycol. Talanta 77:262–268PubMedCrossRefGoogle Scholar
  11. Costanzo MC, Fox TD (1988) Transformation of yeast by agitation with glass beads. Genetics 120:667–670PubMedPubMedCentralGoogle Scholar
  12. Delorme E (1989) Transformation of Saccharomyces cerevisiae by electroporation. Appl Environ Microbiol 55:2242–2246PubMedPubMedCentralGoogle Scholar
  13. Denamur E, Lecointre G, Darlu P, Tenaillon O, Acquviva C, Sayada C, Sunjevaric I, Rothstein R, Elion J, Taddei F, Radman M, Matic I (2000) Evolutionary implications of the frequent horizontal transfer of mismatch repair genes. Cell 103:711–721PubMedCrossRefGoogle Scholar
  14. Durand J, Birdsell J, Wills C (1993) Pleiotropic effects of heterozygosity at the mating-type locus of the yeast Saccharomyces cerevisiae on repair, recombination and transformation. Mutat Res 290:239–247PubMedCrossRefGoogle Scholar
  15. Elkhaimi M, Kaadige MR, Kamath D, Jackson JC, Biliran H Jr, Lopes JM (2000) Combinatorial regulation of phospholipid biosynthetic gene expression by the UME6, SIN3 and RPD3 genes. Nucleic Acids Res 28:3160–3167PubMedCrossRefPubMedCentralGoogle Scholar
  16. Elouahabi A, Ruysschaert JM (2005) Formation and intracellular trafficking of lipoplexes and polyplexes. Mol Ther 11:336–347PubMedCrossRefGoogle Scholar
  17. Fitzpatrick DA (2012) Horizontal gene transfer in fungi. FEMS Microbiol Lett 329:1–8PubMedCrossRefGoogle Scholar
  18. Gallego C, Casas C, Herrero E (1993) Increased transformation levels in intact cells of Saccharomyces cerevisiae aculeacin A-resistant mutants. Yeast 9:523–526PubMedCrossRefGoogle Scholar
  19. Gerbaud C, Fournier P, Blanc H, Aigle M, Heslot H, Guerineau M (1979) High frequency of yeast transformation by plasmids carrying part or entire 2-μm yeast plasmid. Gene 5:233–253PubMedCrossRefGoogle Scholar
  20. Gietz RD, Schiestl RH (2007a) Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:1–4PubMedCrossRefGoogle Scholar
  21. Gietz RD, Schiestl RH (2007b) Microtiter plate transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:5–8PubMedCrossRefGoogle Scholar
  22. Gietz RD, Schiestl RH (2007c) High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:31–34PubMedCrossRefGoogle Scholar
  23. Gietz RD, Schiestl RH (2007d) Quick and easy yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:35–37PubMedCrossRefGoogle Scholar
  24. Gietz RD, Schiestl RH (2007e) Large-scale high-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:38–41PubMedCrossRefGoogle Scholar
  25. Gietz RD, Woods RA (2001) Genetic transformation of yeast. Biotechniques 30:816–820, 822–826, 828 passimGoogle Scholar
  26. Gietz RD, Schiestl RH, Willems AR, Woods RA (1995) Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360PubMedCrossRefGoogle Scholar
  27. Griffith F (1928) The significance of pneumococcal types. J Hyg (Lond) 27:113–159CrossRefGoogle Scholar
  28. Hayama Y, Fukuda Y, Kawai S, Hashimoto W, Murata K (2002) Extremely simple, rapid and highly efficient transformation method for the yeast Saccharomyces cerevisiae using glutathione and early log phase cells. J Biosci Bioeng 94:166–171PubMedCrossRefGoogle Scholar
  29. Heinemann JA, Sprague GF Jr (1989) Bacterial conjugative plasmids mobilize DNA transfer between bacteria and yeast. Nature 340:205–209PubMedCrossRefGoogle Scholar
  30. Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci U S A 75:1929–1933PubMedCrossRefPubMedCentralGoogle Scholar
  31. Hooykaas PJ, den Dulk-Ras A, Bundock P, Soltani J, van Attikum H, van Heusden GP (2006) Yeast (Saccharomyces cerevisiae). Methods Mol Biol 344:465–473PubMedGoogle Scholar
  32. Hsiao CL, Carbon J (1979) High-frequency transformation of yeast by plasmids containing the cloned yeast ARG4 gene. Proc Natl Acad Sci U S A 76:3829–3833PubMedCrossRefPubMedCentralGoogle Scholar
  33. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168PubMedPubMedCentralGoogle Scholar
  34. Jaspersen SL, Ghosh S (2012) Nuclear envelope insertion of spindle pole bodies and nuclear pore complexes. Nucleus 3:226–236PubMedCrossRefPubMedCentralGoogle Scholar
  35. Johnston J, Hilger F, Mortimer R (1981) Variation in frequency of transformation by plasmid YRp7 in Saccharomyces cerevisiae. Gene 16:325–329PubMedCrossRefGoogle Scholar
  36. Johnston SA, Anziano PQ, Shark K, Sanford JC, Butow RA (1988) Mitochondrial transformation in yeast by bombardment with microprojectiles. Science 240: 1538–1541PubMedCrossRefGoogle Scholar
  37. Karube I, Tamiya E, Matsuoka H (1985) Transformation of Saccharomyces cerevisiae spheroplasts by high electric pulse. FEBS Lett 182:90–94CrossRefGoogle Scholar
  38. Kawai S, Pham T, Nguyen HT, Nankai H, Utsumi T, Fukuda Y, Murata K (2004) Molecular insights on DNA delivery into Saccharomyces cerevisiae. Biochem Biophys Res Commun 317:100–107PubMedCrossRefGoogle Scholar
  39. Kawai S, Phan TA, Kono E, Harada K, Okai C, Fukusaki E, Murata K (2009) Transcriptional and metabolic response in yeast Saccharomyces cerevisiae cells during polyethylene glycol-dependent transformation. J Basic Microbiol 49:73–81PubMedCrossRefGoogle Scholar
  40. Kawai S, Hashimoto W, Murata K (2010) Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism. Bioeng Bugs 1:395–403PubMedCrossRefPubMedCentralGoogle Scholar
  41. Keszenman-Pereyra D, Hieda K (1988) A colony procedure for transformation of Saccharomyces cerevisiae. Curr Genet 13:21–23PubMedCrossRefGoogle Scholar
  42. Khalil IA, Kogure K, Akita H, Harashima H (2006) Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery. Pharmacol Rev 58: 32–45PubMedCrossRefGoogle Scholar
  43. Khan NC, Sen S (1974) Genetic transformation in yeasts. J Gen Microbiol 83:237–250PubMedCrossRefGoogle Scholar
  44. Klebe RJ, Harriss JV, Sharp ZD, Douglas M (1983) A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast. Gene 25:333–341PubMedCrossRefGoogle Scholar
  45. Kohiyama M, Hiraga S, Matic I, Radman M (2003) Bacterial sex: playing voyeurs 50 years later. Science 301:802–803PubMedCrossRefGoogle Scholar
  46. Krüger NJ, Stingl K (2011) Two steps away from novelty—principles of bacterial DNA uptake. Mol Microbiol 80:860–867PubMedCrossRefGoogle Scholar
  47. Lacroix B, Tzfira T, Vainstein A, Citovsky V (2006) A case of promiscuity: Agrobacterium’s endless hunt for new partners. Trends Genet 22:29–37PubMedCrossRefGoogle Scholar
  48. Manivasakam P, Schiestl RH (1993) High efficiency transformation of Saccharomyces cerevisiae by electroporation. Nucleic Acids Res 21:4414–4415PubMedCrossRefPubMedCentralGoogle Scholar
  49. Maynard Smith J, Dowson CG, Spratt BG (1991) Localized sex in bacteria. Nature 349:29–31CrossRefGoogle Scholar
  50. Mellman I, Fuchs R, Helenius A (1986) Acidification of the endocytotic and exocytic pathways. Annu Rev Biochem 55:663–700PubMedCrossRefGoogle Scholar
  51. Mitrikeski PT (2013) Yeast competence for exogenous DNA uptake: towards understanding its genetic component. Antonie Van Leeuwenhoek 103:1181–1207PubMedCrossRefGoogle Scholar
  52. Neukamm B, Stahl U, Lang C (2002) Endocytosis is involved in DNA uptake in yeast. Biochim Biophys Acta 1572:67–76PubMedCrossRefGoogle Scholar
  53. Neumann E, Kakorin S, Tsoneva I, Nikolova B, Tomov T (1996) Calcium-mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys J 71:868–877PubMedCrossRefPubMedCentralGoogle Scholar
  54. Nevoigt E, Fassbender A, Stahl U (2000) Cells of the yeast Saccharomyces cerevisiae are transformable by DNA under non-artificial conditions. Yeast 16: 1107–1110PubMedCrossRefGoogle Scholar
  55. Nishikawa M, Suzuki K, Yoshida K (1990) Structural and functional stability of IncP plasmids during stepwise transmission by trans-kingdom mating: promiscuous conjugation of Escherichia coli and Saccharomyces cerevisiae. Jpn J Genet 65:323–334PubMedCrossRefGoogle Scholar
  56. Orr-Weaver TL, Szostak JW, Rothstein RJ (1983) Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol 101:228–245PubMedCrossRefGoogle Scholar
  57. Pham TA, Kawai S, Kono E, Murata K (2011a) The role of cell wall revealed by the visualization of Saccharomyces cerevisiae transformation. Curr Microbiol 62:956–961PubMedCrossRefGoogle Scholar
  58. Pham TA, Kawai S, Murata K (2011b) Visualization of the synergistic effect of lithium acetate and single-stranded carrier DNA on Saccharomyces cerevisiae transformation. Curr Genet 57:233–239PubMedCrossRefGoogle Scholar
  59. Piers KL, Heath JD, Liang X, Stephens KM, Nester EW (1996) Agrobacterium tumefaciens-mediated transformation of yeast. Proc Natl Acad Sci U S A 93: 1613–1618PubMedCrossRefPubMedCentralGoogle Scholar
  60. Riechers SP, Stahl U, Lang C (2009) Defects in intracellular trafficking and endocytotic/vacuolar acidification determine the efficiency of endocytotic DNA uptake in yeast. J Cell Biochem 106:327–336PubMedCrossRefGoogle Scholar
  61. Riechers SP, Stahl U, Lang C (2010) Endocytotic uptake of fluorescence labelled DNA in yeast. J Basic Microbiol 50:83–89PubMedCrossRefGoogle Scholar
  62. Robertson AS, Smythe E, Ayscough KR (2009) Functions of actin in endocytosis. Cell Mol Life Sci 66:2049–2065PubMedCrossRefGoogle Scholar
  63. Sawasaki Y, Inomata K, Yoshida K (1996) Trans-kingdom conjugation between Agrobacterium tumefaciens and Saccharomyces cerevisiae, a bacterium and a yeast. Plant Cell Physiol 37:103–106PubMedCrossRefGoogle Scholar
  64. Schiestl RH, Gietz RD (1989) High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16:339–346PubMedCrossRefGoogle Scholar
  65. Soltani J, van Heusden GP, Hooykaas PJ (2009) Deletion of host histone acetyltransferases and deacetylases strongly affects Agrobacterium-mediated transformation of Saccharomyces cerevisiae. FEMS Microbiol Lett 298:228–233PubMedCrossRefGoogle Scholar
  66. Stateva LI, Oliver SG, Trueman LJ, Venkov PV (1991) Cloning and characterization of a gene which determines osmotic stability in Saccharomyces cerevisiae. Mol Cell Biol 11:4235–4243PubMedPubMedCentralGoogle Scholar
  67. Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A 76:1035–1039PubMedCrossRefPubMedCentralGoogle Scholar
  68. Tomlin GC, Hamilton GE, Gardner DCJ, Walmsley RM, Stateva LI, Oliver SG (2000) Suppression of sorbitol dependence in a strain bearing a mutation in the SRB1/PSA1/VIG9 gene encoding GDP-mannose pyrophosphorylase by PDE2 overexpression suggests a role for the Ras/cAMP signal-transduction pathway in the control of yeast cell-wall biogenesis. Microbiology 146:2133–2146PubMedGoogle Scholar
  69. Tsuchiya E, Shakuto S, Miyakawa T, Fukui S (1988) Characterization of a DNA uptake reaction through the nuclear membrane of isolated yeast nuclei. J Bacteriol 170:547–551PubMedPubMedCentralGoogle Scholar
  70. Vandamme J, Castermans D, Thevelein JM (2012) Molecular mechanisms of feedback inhibition of protein kinase A on intracellular cAMP accumulation. Cell Signal 24:1610–1618PubMedCrossRefGoogle Scholar
  71. Wang H, Clark I, Nicholson PR, Herskowitz I, Stillman DJ (1990) The Saccharomyces cerevisiae SIN3 gene, a negative regulator of HO, contains four paired amphipathic helix motifs. Mol Cell Biol 10:5927–5936PubMedPubMedCentralGoogle Scholar
  72. Wattiaux R, Laurent N, Wattiaux-De Coninck S, Jadot M (2000) Endosomes, lysosomes: their implication in gene transfer. Adv Drug Deliv Rev 41:201–208PubMedCrossRefGoogle Scholar
  73. Zheng HZ, Liu HH, Chen SX, Lu ZX, Zhang ZL, Pang DW, Xie ZX, Shen P (2005) Yeast transformation process studied by fluorescence labeling technique. Bioconjug Chem 16:250–254PubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Laboratory for Evolutionary Genetics, Division of Molecular BiologyRuđer Bošković InstituteZagrebCroatia
  2. 2.Institute for Research and Development of Sustainable Ecosystems, FSB-CTTZagrebCroatia

Personalised recommendations