Abstract
The antimutator phenotype, reportedly conferred by disruption of the Saccharomyces cerevisiae DDR48 gene, was suggested to affect only a specific spontaneous mutational pathway. We attempted to identify the types of mutation that are DDR48-dependent by determining the specificity of the ddr48 antimutator. However, disruption of DDR48 did not decrease the rates of spontaneous forward mutation in a plasmid-borne copy of the yeast SUP4-o gene, the reversion or suppression of the lys2–1 allele, or forward mutation at the CAN1 locus. Interestingly, the latter gene had been reported previously to be subject to the antimutator effect. DNA sequence analysis of spontaneous SUP4-o mutations arising in DDR48 and ddr48 backgrounds provided no evidence for a reduction in the rates of individual mutational classes. Thus, we were unable to verify that disruption of DDR48 causes an antimutator phenotype.
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Boeke J, LaCroute F, Fink G (1984) A positive selection for mutants lacking orotidine-5′ phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet 197: 345–346
Donahue TF, Farabaugh PJ, Fink GR (1981) Suppressible four-base-glycine and proline codons in yeast. Science 212: 455–457
Drake JW (1991) A constant rate of spontaneous mutation in DNA-based microbes. Proc Natl Acad Sci USA 88: 7160–7164
Gietz D, Sugino A (1988) New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74: 527–534
Gietz D, St. Jean A, Woods RA, Schiest RH (1992) Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20: 1425
Impellizzeri KJ, Anderson B, Burgers PMJ (1991) The spectrum of spontaneous mutations in a Saccharomyces cerevisiae uracil-DNA-glycosylase mutant limits the function of this enzyme to cytosine deamination repair. J Bacteriol 173: 6807–6810
Kang X, Yadao F, Gietz RD, Kunz BA (1992) Elimination of the yeast RAD6 ubiquitin conjugase enhances base-pair transitions and G·C→T·A transversions as well as transposition of the Ty element: implications for the control of spontaneous mutagenesis. Genetics 130: 285–294
Kohalmi L, Kunz BA (1992) In vitro mutagenesis of the yeast SUP4-o gene to identify all substitutions that can be detected in vivo with the SUP4-o system. Environ Mol Mutagenesis 19:282–287
Kohalmi SE, Glattke M, McIntosh EM, Kunz BA (1991) Mutational specificity of DNA precursor-pool imbalances in yeast arising from deoxycytidylate deaminase deficiency or treatment with thymidylate. J Mol Biol 220: 933–946
Kunz BA, Pierce MK, Mis JRA, Giroux CN (1987) DNA sequence analysis of the muational specificity of u.v. light in the SUP4-o gene of yeast. Mutagenesis 2: 445–453
Kunz BA, Peters MG, Kohalmi SE, Armstrong JD, Glattke M, Badiani K (1989) Disruption of the RAD52 gene alters the spectrum of spontaneous mutations in Saccharomyces cerevisiae. Genetics 122: 535–542
Kunz BA, Kohalmi L, Kang X, Magnusson KA (1990) Specificity of the mutator effect caused by disruption of the RAD1 excision repair gene of Saccharomyces cerevisiae. J Bacteriol 172: 3009–3014
Kunz BA, Kang X, Kohalmi L (1991) The yest rad18 mutator specificially increases G·C→T·A transversions without reducing correction of G-A or C-T mismatches to G·C pairs. Mol Cell Biol 11: 218–225
Kunz BA, Henson ES, Roche H, Ramotar D, Nunoshiba T, Demple B (1994) Specificity of the mutator caused by deletion of the yeast structural gene (APN1) for the major apurinic endonuclease. Proc Natl Acad Sci USA 91: 8165–8169
McClanahan T, McEntee K (1986) DNA damage and heat shock dually regulate genes in Saccharomyces cerevisiae. Mol Cell Biol 6: 90–96
Mis JRA, Kunz BA (1990) Analysis of mutations induced in the SUP4-o gene of Saccharomyces cerevisiae by cis-diamine dichloroplatinum (II). Carcinogenesis 11: 633–638
Newlon CS (1988) Yeast chromosome replication and segregation. Microbiol Rev 52: 568–601
Pierce MK, Giroux CN, Kunz BA (1987) Development of a yeast system to assay mutational specificity. Mutat Res 182: 65–74
Roche H, Gietz RD, Kunz BA (1994) Specificity of the yeast rev3Δ antimutator and REV3 dependency of the mutator resulting from a defect (rad1Δ) in nucleotide excision repair. Genetics 137: 637–646
Sherman F (1982) Suppression in yeast. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces. Metabolism and gene expression. Cold Spring Harbor Laboratory. Cold Spring Harbor, New York, pp 463–486
Sokal RR, Rohlf FJ (1969) Biometry. WH Freeman, San Francisco, California
Treger JM, McEntee K (1990) Structure of the DNA damage-inducible gene DDR48 and evidence for its role in mutagenesis in Saccharomyces cerevisiae. Mol Cell Biol 10: 3174–3184
Wang SS, Hopper AK (1988) Isolation of a yeast gene involved in species-specific pre-tRNA processing. Mol Cell Biol 8: 5140–5149
Yang Y, Kunz BA (1994) Naturally-occurring single nucleotide loops are corrected more efficiently than base mismatches in yeast. Environ Mol Mutagenesis 23 suppl:75
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Roche, H., Ramachandran, K. & Kunz, B.A. Failure to detect an antimutator phenotype following disruption of the Saccharomyces cerevisiae DDR48 gene. Curr Genet 27, 496–500 (1995). https://doi.org/10.1007/BF00314438
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DOI: https://doi.org/10.1007/BF00314438