Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The Escherichia coli recA gene increases resistance of the yeast Saccharomyces cerevisiae to ionizing and ultraviolet radiation


The Escherichia coli recA protein coding region was ligated into an extrachromosomally replicating yeast expression vector downstream of the yeast alcohol dehydrogenase promoter region to produce plasmid pADHrecA. Transformation of the wild-type yeast strains YNN-27 and 7799-4B, as well as the recombination-deficient rad52-t C5-6 mutant, with this shuttle plasmid resulted in the expression of the bacterial 38 kDa RecA protein in exponential phase cells. The wild-type YNN27 and 7799-4B transformants expressing the bacterial recA gene showed increased resistance to the toxic effects of both ionizing and ultraviolet radiation. RecA moderately stimulated the UV-induced mutagenic response of 7799-4B cells. Transformation of the rad52-t mutant with plasmid pADHrecA did not result in the complementation of sensitivity to ionizing radiation. Thus, the RecA protein endows the yeast cells with additional activities, which were shown to be error-prone and dependent on the RAD52 gene.

This is a preview of subscription content, log in to check access.


  1. Aguilera A, Klein HL (1989) Yeast intrachromosomal recombination: Long gene conversion tracts are preferentially associated with reciprocal exchange and require the RAD1 and RAD3 products. Genetics 123:683–694

  2. Andrade HHR, Marques EK, Schenberg ACG, Henriques JAP (1989) The PSO4 gene is responsible for an error-prone recombinational DNA repair pathway in Saccharomyces cerevisiae. Mot Gen Genet 217:419–426

  3. Angulo JF, Schwencke J, Moreau PL, Moustachi E, Devoret R (1985) A yeast protein analogous to Escherichia coli RecA protein whose cellular level is enhanced after UV-irradiation. Mot Gen Genet 201:20–24

  4. Angulo JF, Moreau PL, Maunoury R, Laporte J, Hill AM, Bertolotti R, Devoret R (1989) KIN, a mammalian nuclear protein immunologically related to E. coli RecA protein. Mutat Res 217:123–134

  5. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254

  6. Brennand J, Margison GP (1986) Reduction of the toxicity and mutagenicity of alkylation agents in mammalian cells harbouring the E. coli alkyltransferase gene. Proc Natl Acad Sci USA 83:6292–6296

  7. Brozmanova J, Iljina TP, Saenko AS, Sedliaková M (1981) Inducible repair of DNA double strand breaks in Escherichia coli. Studia Biophys 85:195–201

  8. Brozmanova J, Kleibl K, Vlčková V, Škorvaga M, Cernakova L, Margison GP (1990) Expression of the E. coli ada gene in yeast protects against the toxic and mutagenic effects of N-methyl-Nnitro-N-nitrosoguanidine. Nucleic Acids Res 18:331–335

  9. Brunborg G, Williamson DH (1978) The relevance of the nucleardivision cycle to radiosensitivity in yeast. Mol Gen Genet 162:277–286

  10. Brunborg G, Resnick MA, Williamson DH (1980) Cell-cycle-specific repair of DNA double-strand breaks in Saccharomyces cerevisiae. Radiat Res 82:547–558

  11. Cernakova L, Piršel M, Kleibl K, Brozmanova J (1989) Plasmid construction for transfer of the recA gene from E. coli to S. cerevisiae using Ba131 digestion. Biológia Bratislava 44:297–305

  12. Chen DS, Bernstein H (1988) Yeast gene RAD52 can substitute for phage T4 gene 46 and 47 in earring out recombination and DNA repair. Proc Natl Acad Sci USA 85:6821–6825

  13. Chenevert JA, Naumovski L, Schultz RA, Friedberg EC (1986) Partial complementation of the UV sensitivity of E. coli and yeast excision repair mutants by the cloned den V gene of bacteriophage T4. Mol Gen Genet 203:163–171

  14. Cole GH, Schild D, Lovett ST, Mortimer RK (1987) Regulation of RAD54- and RAD52-lacZ gene fusions in Saccharomyces cerevisiae in response to DNA damage. Mol Cell Biol 7:1078–1084

  15. Cox MM, Lehman IR (1987) Enzymes of general recombination. Annu Rev Biochem 56:229–262

  16. Dykstra CC, Mittis J, Resnick MA, Sugino A (1988) ATP-dependent strand transfer catalyzed by proteins from the yeast Saccharomycas cerevisiae. In: Friedberg EC, Hanawalt PC (eds) Mechanisms and Consequences of DNA Damage Processing. Alan R. Liss, New York, pp 231–235

  17. Eitner G, Solonin AS, Tanyashin VI (1981) Cloning of a recA-like gene of Proteus mirabilis. Gene 14:301–308

  18. Elledge JS, Davis RW (1987) Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene. Mol Cell Biol 7:2783–2793

  19. Fabre F (1973) The role of repair mechanisms in the variations of ultraviolet and gamma-radiation sensitivity during the cell cycle of Schizosaccharomyces pombe. Radiat Res 56:528–539

  20. Fink PW, Brough CL, Emmerson PT (1986) Molecular cloning of a recA-like gene of Methylophilus methyltrophus and identification of its product. Gene 44:47–53

  21. Game JC, Mortimer RK (1974) A genetic study of X-ray sensitive mutants in yeast. Mutat Res 24:281–292

  22. Goodman HJK, Parker JR, Southern JA, Woods DR (1987) Cloning and expression in Escherichia coli of a recA-like gene from Bacteroides fragilis. Gene 58:265–271

  23. Harries BD, Rickwood D (1981) Gel electrophoresis of proteins. IRL Press, Washington, DC

  24. Ho KSY (1975) Induction of DNA double strand breaks by X-ray in a radiation sensitive strain of the yeast Saccharomyces cerevisae. Mutat Res 30:327–334

  25. Horii T, Ogawa T, Ogawa H (1980) Organization of the recA gene of Escherichia coli. Proc Natl Acad Sci USA 77:313–317

  26. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact cells treated with alkali cations. J Bacteriol 153:163–168

  27. Iwaki T, Iwaki AK, Liem RKH, Goodman JE (1989) αβ-Crystallin is expressed in non-leuticular tissues and accumulates in Alexander's disease brain. Cell 57:71–78

  28. Klein HL (1988) Different types of recombination events are controlled by the RAD1 and RAD52 genes of Saccharomyces cerevisiae. Genetics 120:367–377

  29. Kmiec E, Holloman WK (1982) Homologous pairing of DNA molecules promoted by a protein from Ustilago. Cell 29:367–374

  30. Kokjohn TA, Miller RV (1985) Molecular cloning and characterization of the recA gene of Pseudomonas aeruginosa PAO. J Bacteriol 163:568–572

  31. Krasin F, Hutchinson F (1977) Repair of DNA double-strand breaks in Escherichia coli which requires recA function and the presence of a duplicate genome. J Mol Biol 116:81–98

  32. Krasin F, Hutchinson F (1981) Repair of DNA-double strand breaks in Escherichia coli requires synthesis of proteins that can be induced by UV light. Proc Natl Acad Sci USA 78:3450–3453

  33. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

  34. Langeveld SA, Yasui A, Eker APM (1985) Expression of an Escherichia coli phr gene in the yeast Saccharomyces cerevisiae. Mol Gen Genet 199:396–400

  35. Langguth EN, Beam CA (1973) Repair mechanisms and cell cycle dependent variations in X-ray sensitivity of diploid yeast. Radiat Res 53:226–234

  36. Lovett CM, Roberts JW (1985) Purification of a RecA protein from Bacillus subtilis. J Biol Chem 260:3305–3313

  37. Lu C, Scheuermann RH, Harrison E (1986) Capacity of RecA protein to bind preferentially to UV lesions and inhibit the editing subunit of DNA polymerase III: A possible mechanism for SOS-induced targeted mutagenesis. Proc Natl Acad Sci USA 83:619–623

  38. Madura K, Prakash S (1986) Nucleotide sequence, transcript mapping and regulation of the RAD2 gene of Saccharomyces cerevisiae. J Bacteriol 166:914–923

  39. Malone RE, Esposito RE (1980) The RAD52 gene product is required for homothalic interconversion of mating types and spontaneous mitotic recombination. Proc Natl Acad Sci USA 77:503–507

  40. Newlon CS (1988) Yeast chromosome replication and segregation. Microbiol Rev 52:568–601

  41. Pierre A, Paoletti C (1983) Purification and characterization of RecA protein from Salmonella typhimurium. J Biol Chem 258:2870–2874

  42. Resnick MA (1969) Genetic control of radiation sensitivity in Saccharomyces cerevisiae. Genetics 62:519–531

  43. Resnick MA (1979) The induction of molecular and genetic recombination in eukaryotic cells. Adv Radiat Biol 8:175–217

  44. Resnick MA, Martin P (1976) The repair of double-strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control. Mol Gen Genet 143:119–129

  45. Robinson GW, Nicolet CM, Kalainov D, Friedberg EC (1986) A yeast excision-repair gene is inducible by DNA damaging agents. Proc Natl Acad Sci USA 83:1842–1846

  46. Saeki T, Machida I, Nakai S (1980) Genetic control of diploid recovery after γ-irradiation in the yeast Saccharomyces cerevisiae. Mutat Res 73:251–265

  47. Samson L, Derfler B, Waldstein EA (1986) Suppression of human DNA alkylation DNA repair defects by Escherichia coli DNA repair genes. Proc Natl Acad Sci USA 83:5607–5610

  48. Sancar A, Stachelek GC, Konigsberg W, Rupp WD (1980) Sequence of the recA gene and protein. Proc Natl Acad Sci USA 77:2611–2615

  49. Sargentini NJ, Smith KC (1986) Characterization and quantitation of DNA strand breaks requiring recA-dependent repair in X-irradiated Escherichia coli. Radiat Res 105:180–186

  50. Schiestl A, Prakash S (1988) RAD1, an excision repair gene of Saccharomyces cerevisiae is also involved in recombination. Mol Cell Biol 8:3619–3626

  51. Schiestl A, Prakash S (1990) RAD10, an excision repair gene of Saccharomyces cerevisiae is involved in the RAD1 pathway of mitotic recombination. Mol Cell Biol 10:2485–2491

  52. Shibata T, Cunningham RP, Radding CM (1981) Homologous pairing in genetic recombination, purification and characterization of Escherichia coli RecA protein. J Biol Chem 256:7557–7564

  53. Shinagawa H, Iwasaki H, Kato T, Nakata A (1988) RecA proteindependent cleavage of UmuD protein and SOS mutagenesis. Proc Natl Acad Sci USA 85:1806–1810

  54. Smith KC, Wang TW (1989) recA-Dependent DNA repair processes. Bio-essays 10:12–16

  55. Symington L (1990) Double strand-break repair and recombination catalyzed by a nuclear extract from yeast. Book of Abstracts, RecA and Related Proteins. Saclay, France Sept 17–21, Abstract No 44

  56. Szostak JW, Orr-Weaver TL, Rothstein RJ (1983) The double strand-break repair model for recombination. Cell 33:25–35

  57. Terry Y, Chow K, Resnick MA (1987) Purification and characterization of endo-exonuclease from Saccharomyces cerevisiae that is influenced by the RAD52 gene. J Biol Chem 262:17659–17667

  58. Terry Y, Chow K, Resnick MA (1988) An endo-exonuclease activity of yeast that requires a functional RAD52 gene. Mol Gen Genet 211:41–48

  59. 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 76:4350–4354

  60. Uhlin BE, Clark AJ (1981) Overproduction of the Escherichia coli RecA protein without stimulation of its proteolytic activity. J Bacteriol 148:386–390

  61. Valerie K, Fronko G, Henderson EE, Riel JK (1986) Expression of the den V gene of coliphage T4 in UV sensitive rad mutants of Saccharomyces cerevisiae. Mol Cell Biol 6:3559–3562

  62. Valerie K, Green AP, Riel JK, Henderson EE (1987) Transient and stable complementation of ultraviolet repair in XP cells by the denV gene of bacteriophage T4. Cancer Res 47:2967–2971

  63. Walker GC (1984) Mutagenesis and inducible responses to DNA damage in Escherichia coli. Microbiol Rev 48:60–93

  64. West SC, Cassuto E, Howard-Flanders P (1981) Mechanism of E. coli RecA protein directed strand exchange in post-replication repair of DNA. Nature 294:659–662

Download references

Author information

Correspondence to Jela Brozmanová.

Additional information

Communicated by R. Devoret

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Brozmanová, J., Černáková, Ľ., Vlčková, V. et al. The Escherichia coli recA gene increases resistance of the yeast Saccharomyces cerevisiae to ionizing and ultraviolet radiation. Molec. Gen. Genet. 227, 473–480 (1991).

Download citation

Key words

  • Saccharomyces cerevisiae
  • recA gene expression
  • Resistance to ionizing and ultraviolet radiation