Animal cells and systems

The Korean Society for Integrative Biology (한국통합생물학회)
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Functional Analysis of RAD4 Gene Required for Nucleotide Excision Repair of UV-induced DNA Damage in Saccharomyces cerevisiae

  • Park, Sang Dai (Faculty of Biological Sciences, Seoul National University) ;
  • Park, In Soon (Department of Life Science, College of Natural Sciences, Silla University)
  • Published : 2002.12.01
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Abstract

The RAD4 gene is essential for nucleotide excision repair in Saccharomyces cerevisiae. It has been known that the deduced amino acid sequence of Rad4 protein contains three DNA-dependent ATPase/helicase motifs. To determine the biochemical activities and functional role of RAD4 the Rad4 protein was expressed and purified. Immunoblot analysis showed a specific band of 21 kDa, which was well-matched with the size of open reading frame of the RAD4 gene. The purified Rad4 protein had no detectable helicase activity. However, the protein could interact with double stranded oligonucleotides, as judged by mobility shift assay. This result suggests that the Rad4 protein is a DNA binding protein.

Keywords

References

  1. Auble DT, Hansen KE, Mueller CG, Lane WS, Thorner J, and Hahn S (1994) Mot1, a global repressor of RNA polymerase II transcription, inhibits TBP binding to DNA by ATP-dependent mechanism. Genes & Dev 8: 1920-1934 https://doi.org/10.1101/gad.8.16.1920
  2. Boothmann DA, Meyers M, Fukunaga N, and Lee SW (1993) Isolation of X-ray-inducible transcripts from radioresistant human melanoma cells. Proc Natl Acid Sci USA 90: 7200-7204 https://doi.org/10.1073/pnas.90.15.7200
  3. Choi IS, Kim JB, and Park SD (1990) Nucleotide sequence of RAD4 gene of Saccharomyces cerevisiae that can be propagated in Escherichia coli without inactivation. Nucleic Acids Res 18: 7137 https://doi.org/10.1093/nar/18.23.7137
  4. Choi IS, Kim JB, Hong SH, and Park SD (1991) A gene in Schizosaccharomyces pombe analogous to the RAD4 gene of Saccharomyces cerevisiae. FEMS Microbiol Lett 77 : 97-100 https://doi.org/10.1111/j.1574-6968.1991.tb04328.x
  5. Choi IS, Kim JB, Jeon SH, and Park SD (1993) Expression of RAD4 gene of Saccharomyces cerevisiae that can be propagated in Escherichia coli without inactivation. Biochem Biophy Res Commun 193: 91-197 https://doi.org/10.1006/bbrc.1993.1608
  6. Choi IS (2001) Isolation and characterization of UV-inducible genes in eukaryotic cells. Korean J Life Sci 11-1: 52-56
  7. Cote J, Quinn J, Workman JL, and Peterson CL (1994) Stimulation of Gal4 derivative binding to nucleosomal DNA by the yeast SW/SNF complex. Science 265: 53-60 https://doi.org/10.1126/science.8016655
  8. Delmas V, Stokes DG, and Perry RP (1993) A mammalian DNA-binding protein that contains a chromodomain and an SNF2/SW12-like helicase domain. Proc Natl Acad Sci USA 90: 2414-2418 https://doi.org/10.1073/pnas.90.6.2414
  9. Fenech M, Car AM, Murray J, Watts FZ, and Lehmann AR (1991) Cloning and characterization of the RAD4 gene of Schizosaccharomyces pombe; a gene showing short regions of sequence similarity to the human ERCC1 gene. Nucleic Acids Res 19-24: 6737-6741 https://doi.org/10.1093/nar/19.24.6737
  10. Fleer R, Nicolet CM, Pure GA, and Friedberg EC (1987) RAD4 gene of Saccharomyces cerevisiae: molecular cloning and partial characterization of a gene that is inactivated in Escherichia coli. Mol Cell Biol 7: 1180-1192 https://doi.org/10.1128/MCB.7.3.1180
  11. Friedberg EC (1988) Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbial Rev 52: 70-102
  12. Henikoff S (1993) Transcriptional activator components and poxvirus DNA-dependent ATPase comprise a single family. Trends Biochem Sci 18: 291-292 https://doi.org/10.1016/0968-0004(93)90037-N
  13. Higgns DR, Prakash S, Reynolds P, Polakowska R, Weber S, and Prakash L (1983) Isolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants. Proc Nat Acad Sci USA 80: 5680-5684 https://doi.org/10.1073/pnas.80.18.5680
  14. Hoeijmakers JHJ and Bootsma D (1990) Molecular genetics of eukaryotic DNA excision repair. Cancer Cells 2: 311-320
  15. Ito H, Fukuda Y, Murata K, and Kimmura A (1983) Transformation of intact yeast cells treated with alkali cations. J. Bacterial 153: 163-168
  16. Jang YK, Jin YH, Kim EM, Fabre F, Hong SH, and Park SD (1993) Molecular cloning of $rhp51^+$ gene in Schizosccharomyces pombe, whose amino acid sequence is highly conserved from prokaryotic RecA to the mammalian Rad51 homolog. Gene 169: 125-130 https://doi.org/10.1016/0378-1119(96)83099-X
  17. Jin YH, Yoo EJ, Jang YK, Kim SH, Lee CG, Seong RH, Hong SH, and Park SD (1998) Purification and characterization of Hrp1, a homolog of mouse CHD1 from fission yeast Schizosaccharomyces pombe. Korean J Biol Sci 2: 539-543
  18. Kim JB, Jeon SH, Choi IS, and Park SD (1994) Overexpressed Rad4 protein required for excision repair of Saccharomyces cerevisiae is toxic to the host Escherichia coli. In Vitro Toxicol 7: 269-275
  19. Lee CG, Chang KA, Kuroda MI, and Hurwitz (1997) The NTPase/helicase activities of Drosophila maleless, an essential factor in dosage compensation. EMBO J 16: 102671-102681 https://doi.org/10.1093/emboj/16.10.2671
  20. Madura K and Prakash S (1990) Transcript levels of the Saccharomyces cerevisiae DNA repair gene RAD23 increase in response to UV light and in meiosis but remain constant in the mitotic cell cycle. Necleic Acids Res. 18: 4737-4742 https://doi.org/10.1093/nar/18.16.4737
  21. Miller RD, Prakash L and Prakash S (1982) Genetic control of excision of Saccharomyces cerevisiae interstrand DNA crosslinks induced by psoralen plus near-UV light. Mol Gen Genet 188: 235-239 https://doi.org/10.1007/BF00332681
  22. Murray JM, Carr AM, Lehmann AR, and Watts FZ (1991) Cloning and characterization of the rad15 gene, a homologue to the S. cerevisiae RAD3 and human ERCC2 gene. Nucleic Acids Res 19: 3525-3531 https://doi.org/10.1093/nar/19.13.3525
  23. Perozz, G and Prakash S (1986) RAD7 gene of Saccharomyses cerevisiae: transcript, nucleotide sequence analysis and functional relationship between the RAD7 and RAD23 gene products. Mol Cell Biol 6: 1497-1507 https://doi.org/10.1128/MCB.6.5.1497
  24. Phipps J, Nasim A, and Miller DR (1985) Recovery, repair, and mutagenesis in Schizosaccharomyces pombe. Adv Genet 23: 1-72 https://doi.org/10.1016/S0065-2660(08)60511-8
  25. Praekelt UM and Macock PA, (1990) HSP12, a new small heat shock gene of Saccharomyces cerevisiae: analysis of structure, regulation and function. Mol Gen Genet 233: 97-106 https://doi.org/10.1007/BF00315801
  26. Reynolds RJ and Friedgerg EC (1981) Molecular mechanisms of pyrimidine dimer excision of ultraviolet-irradiated deoxyribonucleic acid. J Bacterial 146: 692-704
  27. Reynolds PR, Biggar S, Prakash L, and Prakash S (1992) The Shizosaccharomyces pombe rhp3+ gene required for DNA repair and cell viability is functionally interchangeable with the RAD3 gene of Saccharomyces cerevisiae. Nucleic Acids Res 20: 2327-2334 https://doi.org/10.1093/nar/20.9.2327
  28. Sambrook J and Russell DW (2001) Molecular Cloning: a Laboratory Mannual, 3rd Ed. Cold Spring Harbor Laboratory Press, New york
  29. Sanger F, Nicklen S, and Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Nat Acad Sci USA 764: 5463-5467 https://doi.org/10.1073/pnas.74.12.5463
  30. Thompson LH, Mitchell DL, Regan JD, Bouffler SD, Stewart SA, Carrier, WL, Nairn RS, and Johnson RT (1988) CHO mutant UV61 removes photoproducts but not cyclobutane dimers. Mutagenesis 4: 140-146 https://doi.org/10.1093/mutage/4.2.140
  31. Weber CA, Salazar EP, Stewart SA, and Thompson LH (1990) ERCC2: cDNA cloning and molecular characterization of a human nucleotide excision repair gene with high homology to yeast RAD3. EMBO J 9: 1437-1447
  32. Weinert TA and Hartwel LH (1990) Characterization of RAD9 of Saccharomyces cerevisiae and evidence that its function acts posttranslationally in cell cycle arrest after DNA damage. Mol Cell Biol 10: 6554-6564 https://doi.org/10.1128/MCB.10.12.6554
  33. Woodage T, Basrai MA, Baxxevanis AD, Hieter P, and Collins FS (1997) Characterization of the CHD family of proteins. Proc Natl Acad Sci USA 94: 11472-11477 https://doi.org/10.1073/pnas.94.21.11472