Regulation of the RAD2 gene of Saccharomyces cervisiae

Regulation of the DNA damage‐inducible RAD2 gene was investigated in yeast cells transformed with centromeric plasmids containing RAD2‐lacZ fusion constructs. Deletion analysis defined several regions in the 350 bp region upstream of the translational start codon which are required for induction of β‐galactosidase activity. No deletions resulted in constitutivety enhanced expression. We therefore conclude that induction of RAD2 by DNA‐damaging agents is positively regulated. Two domains required for induction have a similar sequence and are located ∼70 and ∼140bp upstream of the major transcriptional start site. Four other sequence domains required for induction contain uninterrupted poly(dA) poly(dT) stretches 9‐13 bp long. Deletion of some of these AT‐rich domains also affects constitutive expression of RAD2. Expression of RAD2 is not cell‐cycle‐regulated in mitotic cells. However, meiosis is accompanied by increased steady‐state levels of RAD2 mRNA in the absence of DNA damage. This enhanced transcription is not dependent on the presence of upstream sequences required for regulation of induction by DNA damage. Increased steady‐state levels of RAD2 mRNA are induced by cycloheximide in asynchronously dividing populations of cells, but not in non‐replicating cells arrested in G1 phase of the cell cycle. Following exposure to u.v. irradiation induction is also dramatically reduced in non‐replicating cells.

[1]  J Cairns,et al.  Cold spring harbor. , 1991, Science.

[2]  R. Mortimer,et al.  Failure to induce a DNA repair gene, RAD54, in Saccharomyces cerevisiae does not affect DNA repair or recombination phenotypes , 1989, Molecular and cellular biology.

[3]  R. Mortimer,et al.  Two DNA repair and recombination genes in Saccharomyces cerevisiae, RAD52 and RAD54, are induced during meiosis , 1989, Molecular and cellular biology.

[4]  E. Winter,et al.  A DNA binding protein that recognizes oligo(dA).oligo(dT) tracts. , 1989, The EMBO journal.

[5]  E. Friedberg,et al.  Nucleotide sequence of the wild-type RAD4 gene of Saccharomyces cerevisiae and characterization of mutant rad4 alleles , 1989, Journal of bacteriology.

[6]  M. Tuite,et al.  Mistranslation induces the heat‐shock response in the yeast Saccharomyces cerevisiae , 1989, Molecular microbiology.

[7]  R. Kornberg,et al.  Activation of yeast RNA polymerase II transcription by a thymidine-rich upstream element in vitro. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[8]  E. Friedberg.  Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. , 1988, Microbiological reviews.

[9]  K. McEntee,et al.  Expression of the yeast UB14 gene increases in response to DNA-damaging agents and in meiosis , 1988, Molecular and cellular biology.

[10]  R. Morimoto,et al.  Posttranscriptional regulation of hsp70 expression in human cells: effects of heat shock, inhibition of protein synthesis, and adenovirus infection on translation and mRNA stability , 1987, Molecular and cellular biology.

[11]  A. Klug,et al.  The structure of an oligo(dA)·oligo(dT) tract and its biological implications , 1987, Nature.

[12]  K. Struhl,et al.  Distinguishing between mechanisms of eukaryotic transcriptional activation with bacteriophage T7 RNA polymerase , 1987, Cell.

[13]  R. W. Davis,et al.  Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene required for mitotic viability , 1987, Molecular and cellular biology.

[14]  Leland H. Johnston,et al.  The yeast DNA polymerase I transcript is regulated in both the mitotic cell cycle and in meiosis and is also induced after DNA damage , 1987, Nucleic Acids Res..

[15]  Kevin Struhl,et al.  Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast , 1987, Cell.

[16]  S. Lovett,et al.  Regulation of RAD54- and RAD52-lacZ gene fusions in Saccharomyces cerevisiae in response to DNA damage , 1987, Molecular and cellular biology.

[17]  L. Guarente.  Regulatory proteins in yeast. , 1987, Annual review of genetics.

[18]  K. Struhl.  Constitutive and inducible Saccharomyces cerevisiae promoters: evidence for two distinct molecular mechanisms , 1986, Molecular and cellular biology.

[19]  Paul M. Sharp,et al.  Codon usage in yeast: cluster analysis clearly differentiates highly and lowly expressed genes , 1986, Nucleic Acids Res..

[20]  S. Prakash,et al.  Nucleotide sequence, transcript mapping, and regulation of the RAD2 gene of Saccharomyces cerevisiae , 1986, Journal of bacteriology.

[21]  E. Friedberg,et al.  A yeast excision-repair gene is inducible by DNA damaging agents. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Garrels,et al.  Inducible cellular responses to DNA damage in mammalian cells. , 1986, Basic life sciences.

[23]  K. Struhl.  Nucleotide sequence and transcriptional mapping of the yeast pet56-his3-ded1 gene region. , 1985, Nucleic acids research.

[24]  L. Johnston,et al.  The nucleotide sequence of the DNA ligase gene (CDC9) from Saccharomyces cerevisiae: a gene which is cell-cycle regulated and induced in response to DNA damage. , 1985, Nucleic acids research.

[25]  L. Prakash,et al.  Nucleotide sequence of the RAD10 gene of Saccharomyces cerevisiae. , 1985, The EMBO journal.

[26]  K. Struhl.  Naturally occurring poly(dA-dT) sequences are upstream promoter elements for constitutive transcription in yeast. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[27]  D. Beier,et al.  Deletion analysis identifies a region, upstream of the ADH2 gene of Saccharomyces cerevisiae, which is required for ADR1-mediated derepression , 1985, Molecular and cellular biology.

[28]  D. Kaback,et al.  Saccharomyces cerevisiae exhibits a sporulation-specific temporal pattern of transcript accumulation , 1985, Molecular and cellular biology.

[29]  S. Aves,et al.  Cloning, sequencing and transcriptional control of the Schizosaccharomyces pombe cdc10 ‘start’ gene. , 1985, The EMBO journal.

[30]  L. Naumovski,et al.  RAD3 gene of Saccharomyces cerevisiae: nucleotide sequence of wild-type and mutant alleles, transcript mapping, and aspects of gene regulation , 1985, Molecular and cellular biology.

[31]  J. Szostak,et al.  Specific Saccharomyces cerevisiae genes are expressed in response to DNA-damaging agents , 1985, Molecular and cellular biology.

[32]  M. Osley,et al.  Regulation of CDC9, the Saccharomyces cerevisiae gene that encodes DNA ligase , 1985, Molecular and cellular biology.

[33]  E. Friedberg.  Nucleotide excision repair of DNA in eukaryotes: comparisons between human cells and yeast. , 1985, Cancer surveys.

[34]  E. Friedberg,et al.  The RAD2 gene of Saccharomyces cerevisiae: nucleotide sequence and transcript mapping. , 1985, Gene.

[35]  M. Resnick,et al.  DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[36]  T. Mcclanahan,et al.  Specific transcripts are elevated in Saccharomyces cerevisiae in response to DNA damage. , 1984, Molecular and cellular biology.

[37]  E. Friedberg,et al.  Molecular cloning and nucleotide sequence analysis of the Saccharomyces cerevisiae RAD1 gene , 1984, Molecular and cellular biology.

[38]  G. Walker.  Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. , 1984, Microbiological reviews.

[39]  L. Naumovski,et al.  Saccharomyces cerevisiae RAD2 gene: isolation, subcloning, and partial characterization , 1984, Molecular and cellular biology.

[40]  S. Stasiewicz,et al.  Meiotic DNA metabolism in wild-type and excision-deficient yeast following UV exposure. , 1983, Genetics.

[41]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[42]  P. T. Magee,et al.  Isolation of genes expressed preferentially during sporulation in the yeast Saccharomyces cerevisiae. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[43]  J. Bennetzen,et al.  Codon selection in yeast. , 1982, The Journal of biological chemistry.

[44]  L. Prakash,et al.  Incision and postincision steps of pyrimidine dimer removal in excision-defective mutants of Saccharomyces cerevisiae , 1981, Journal of bacteriology.

[45]  E. Friedberg,et al.  Molecular Mechanisms of Pyrimidine Dimer Excision in Saccharomyces cerevisiae: Incision of Ultraviolet-Irradiated Deoxyribonucleic Acid In Vivo , 1981, Journal of bacteriology.

[46]  F. Sanger,et al.  Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. , 1980, Journal of molecular biology.

[47]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[48]  D. Williamson.  Replication of the nuclear genome in yeast does not require concomitant protein synthesis. , 1973, Biochemical and Biophysical Research Communications - BBRC.