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PMID: 2827121 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Molecular characterization of the Saccharomyces cerevisiae dihydrofolate reductase gene (DFR1).

Nucleic acids research ·Vol. 15 ·No. 24 ·1987-12-23 ·Pages 10355-71

Lagosky PA, Taylor GR, Haynes RH

Abstract

The complete nucleotide sequence of a 1957 bp DNA fragment containing the dihydrofolate reductase gene (DFR1) of the yeast Saccharomyces cerevisiae is presented. Within this region a single open reading frame of 633 base pairs was found which is capable of encoding a 211 amino acid residue protein with a calculated Mr of 24,233. The amino acid sequence derived from the yeast DFR1 gene shows limited homology with sequences from both eukaryotic and non-eukaryotic DHFR enzymes. Northern blot hybridization reveals that the mRNA from this gene is a 900 base polyadenylated transcript. Yeast strains containing the cloned DFR1 gene on multicopy number shuttle vector plasmids show dramatically enhanced methotrexate resistance. Consensus DNA sequences responsible for RNA polymerase II interaction and general amino acid control in S. cerevisiae are located within the 5'-noncoding region with respect to the open reading frame. The DNA fragment containing these sequences has been shown to be necessary for DFR1 gene expression in both S. cerevisiae and E. coli.

MeSH Terms
Amino Acid Sequence Base Sequence Chromosome Mapping Cloning, Molecular Codon DNA Restriction Enzymes Escherichia coli/genetics Gene Expression Regulation Genes Genes, Fungal Molecular Sequence Data Saccharomyces cerevisiae/genetics Tetrahydrofolate Dehydrogenase/genetics
Chemicals
Codon Tetrahydrofolate Dehydrogenase DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lagosky P A
Department of Biology, York University, Toronto, Ontario, Canada.
Taylor G R
Haynes R H
References (46)
46 references, click to expand
  1. Calcium-dependent bacteriophage DNA infection.
    J Mol Biol. 1970 Oct 14;53(1):159-62 PMID: 4922220
  2. GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast.
    Cell. 1985 Nov;43(1):177-88 PMID: 3907851
  3. Isolation and properties of a thymidylate-less mutant in Saccharomyces cerevisiae.
    Eur J Biochem. 1975 Aug 15;56(2):533-8 PMID: 1100397
  4. Two forms of serine transhydroxymethylase, one absent in a thymidylate-less mutant in Saccharomyces cerevisiae.
    Eur J Biochem. 1976 Apr 15;64(1):7-13 PMID: 6271
  5. Mitochondrial and cytoplasmic distribution in Saccharmoyces cerevisiae of enzymes involved in folate-coenzyme-mediated one-carbon-group transfer. A genetic and biochemical study of the enzyme deficiencies in mutants tmp3 and ade3.
    Eur J Biochem. 1977 Sep 15;79(1):285-92 PMID: 334546
  6. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.
    Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835-8 PMID: 73185
  7. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  8. Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9.
    Gene. 1977;2(2):75-93 PMID: 344136
  9. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  10. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
    J Bacteriol. 1978 Jun;134(3):1141-56 PMID: 149110
  11. Isolation and characterization of yeast mutants auxotrophic for 2'-deoxythymidine 5'-monophosphate.
    Mol Gen Genet. 1979 Jan 10;168(2):141-51 PMID: 377008
  12. Chimeric plasmids for cloning of deoxyribonucleic acid sequences in Saccharomyces cerevisiae.
    J Bacteriol. 1979 Oct;140(1):73-82 PMID: 387728
  13. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  14. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene.
    Gene. 1979 Dec;8(1):121-33 PMID: 395029
  15. The structure of transposable yeast mating type loci.
    Cell. 1980 Mar;19(3):753-64 PMID: 6244896
  16. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli.
    J Mol Biol. 1980 Apr;138(2):179-207 PMID: 6997493
  17. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase.
    Nucleic Acids Res. 1980 May 24;8(10):2255-74 PMID: 6159575
  18. Induction of mitotic recombination in yeast by starvation for thymine nucleotides.
    Proc Natl Acad Sci U S A. 1980 Oct;77(10):6057-61 PMID: 6449701
  19. Construction of plasmid vectors with unique PstI cloning sites in a signal sequence coding region.
    Gene. 1980 Dec;12(3-4):235-41 PMID: 6265317
  20. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase.
    J Biol Chem. 1982 Mar 25;257(6):3018-25 PMID: 6277922
  21. Codon selection in yeast.
    J Biol Chem. 1982 Mar 25;257(6):3026-31 PMID: 7037777
  22. DNA sequence required for efficient transcription termination in yeast.
    Cell. 1982 Mar;28(3):563-73 PMID: 6280875
  23. Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae.
    Nucleic Acids Res. 1982 Apr 24;10(8):2625-37 PMID: 6281737
  24. Isolation of the thymidylate synthetase gene (TMP1) by complementation in Saccharomyces cerevisiae.
    Mol Cell Biol. 1982 Apr;2(4):437-42 PMID: 6287238
  25. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  26. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.
    Gene. 1982 Oct;19(3):259-68 PMID: 6295879
  27. The isolation, characterization, and sequence of the pyruvate kinase gene of Saccharomyces cerevisiae.
    J Biol Chem. 1983 Feb 25;258(4):2193-201 PMID: 6185493
  28. A short nucleotide sequence required for regulation of HIS4 by the general control system of yeast.
    Cell. 1983 Jan;32(1):89-98 PMID: 6337724
  29. Genetic effects of deoxyribonucleotide pool imbalances.
    Environ Mutagen. 1982;4(6):695-725 PMID: 6761109
  30. Transcription terminates in yeast distal to a control sequence.
    Cell. 1983 Jun;33(2):607-14 PMID: 6305514
  31. Nucleotide sequence of the tcml gene (ribosomal protein L3) of Saccharomyces cerevisiae.
    J Bacteriol. 1983 Jul;155(1):8-14 PMID: 6305925
  32. The nucleotide sequence of the trimethoprim-resistant dihydrofolate reductase gene harbored by Tn7.
    Nucleic Acids Res. 1983 Aug 11;11(15):5147-58 PMID: 6308574
  33. Hybridization of nucleic acids directly in agarose gels.
    Anal Biochem. 1983 Jun;131(2):365-72 PMID: 6193737
  34. A DNA sequence analysis package for the IBM personal computer.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):605-14 PMID: 6546433
  35. Point mutations identify the conserved, intron-contained TACTAAC box as an essential splicing signal sequence in yeast.
    Cell. 1984 Mar;36(3):645-53 PMID: 6365330
  36. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.
    Gene. 1983 Dec;26(1):101-6 PMID: 6323249
  37. The functional human dihydrofolate reductase gene.
    J Biol Chem. 1984 Mar 25;259(6):3933-43 PMID: 6323448
  38. Nucleotide sequence reveals overlap between T4 phage genes encoding dihydrofolate reductase and thymidylate synthase.
    J Biol Chem. 1984 May 25;259(10):6261-6 PMID: 6327673
  39. Schizosaccharomyces pombe and Saccharomyces cerevisiae: a look at yeasts divided.
    Cell. 1986 Jun 20;45(6):781-2 PMID: 3518949
  40. Transcription of genes encoding enzymes involved in DNA synthesis during the cell cycle of Saccharomyces cerevisiae.
    Mol Gen Genet. 1986 Sep;204(3):363-6 PMID: 3020375
  41. Induction of intrachromosomal recombination in yeast by inhibition of thymidylate biosynthesis.
    Genetics. 1986 Oct;114(2):375-92 PMID: 3533714
  42. Molecular characterization of the cell cycle-regulated thymidylate synthase gene of Saccharomyces cerevisiae.
    J Biol Chem. 1987 Apr 15;262(11):5298-307 PMID: 3031048
  43. Cell cycle-dependent expression of thymidylate synthase in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Dec;4(12):2858-64 PMID: 6396509
  44. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  45. Cloned mouse ribonucleotide reductase subunit M1 cDNA reveals amino acid sequence homology with Escherichia coli and herpesvirus ribonucleotide reductases.
    J Biol Chem. 1985 Jun 10;260(11):7015-22 PMID: 2581962
  46. Yeast mutants sensitive to trimethoprim.
    Mutat Res. 1975 May;28(2):175-82 PMID: 1094276
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.24.10355
Published
1987-12-23
Pages
10355-71
Language
English
Region
England
NLM ID
0411011
PMCID
PMC339949
Subset
IM
Databases
GENBANK
Y00887
External Links
PubMed source DOI: 10.1093/nar/15.24.10355 PMC Full Text View journal details
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