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PMID: 2355921 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cloning and analysis of a gene involved in DNA repair and recombination, the rad1 gene of Schizosaccharomyces pombe.

Molecular and cellular biology ·Vol. 10 ·No. 7 ·1990-07-00 ·Pages 3750-60

Sunnerhagen P, Seaton BL, Nasim A, Subramani S

Abstract

We have cloned the rad1 gene of Schizosaccharomyces pombe by complementation of the rad1-1 mutant, which is deficient in DNA repair and recombination. The coding region of the gene is 582 base pairs long and contains no introns. The predicted product is a strongly acidic, 22-kilodalton protein containing 194 amino acid residues. This gene does not exhibit significant homology to any other known repair gene. The major transcription start site is at 27 base pairs upstream of the putative start codon. Insertion mutagenesis revealed that besides the coding region, at least 151 base pairs of 5'-flanking sequence are required for full complementing activity. A strain carrying a null allele of rad1 was constructed and found to have a phenotype closely similar to that of the rad1-1 mutant. Expression in Escherichia coli of the coding region yielded a protein product of a size close to that predicted from the DNA sequence. This product reacted with antibodies raised against a synthetic peptide with a sequence from that predicted for the protein product. We have localized the rad1 gene to NotI fragment E of the S. pombe genome.

MeSH Terms
Amino Acid Sequence Base Sequence Blotting, Southern DNA Repair DNA, Fungal/genetics,isolation & purification Dose-Response Relationship, Radiation Gamma Rays Genes, Viral Genetic Complementation Test Molecular Sequence Data Oligonucleotide Probes RNA, Fungal/genetics,isolation & purification Recombination, Genetic Restriction Mapping Saccharomycetales/genetics Schizosaccharomyces/genetics,radiation effects Ultraviolet Rays
Chemicals
DNA, Fungal Oligonucleotide Probes RNA, Fungal
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Sunnerhagen P
Department of Biology B-022, University of California, San Diego, La Jolla 92093.
Seaton B L
Nasim A
Subramani S
References (44)
44 references, click to expand
  1. Repair-mechanisms and radiation-induced mutations in fission yeast.
    Genetics. 1968 Jul;59(3):327-33 PMID: 5705827
  2. Nuclear pre-mRNA splicing in the fission yeast Schizosaccharomyces pombe strictly requires an intron-contained, conserved sequence element.
    EMBO J. 1987 Jun;6(6):1757-63 PMID: 3649292
  3. The isolation and genetic classification of UV-sensitive mutants of Schizosaccharomyces pombe.
    Mutat Res. 1971 Apr;11(4):361-71 PMID: 5111492
  4. Relation between repair mechanisms and induced mitotic recombination after UV irradiation, in the yeast Schizosaccharomyces pombe. Effects of caffeine.
    Mol Gen Genet. 1972;117(2):153-66 PMID: 5073988
  5. Dark repair inhibitors and pathways for repair of radiation damage in Schizosaccharomyces pombe.
    Mol Gen Genet. 1974;132(1):13-22 PMID: 4417303
  6. Genetic control of radiation sensitivity in Schizosaccharomyces pombe.
    Genetics. 1975 Apr;79(4):573-82 PMID: 166019
  7. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  8. Effect of protein synthesis inhibition on recovery of UV- and gamma-irridated Schizosaccharomyces pombe from repair inhibition by caffeine.
    Mol Gen Genet. 1976 Apr 23;145(1):1-5 PMID: 1272249
  9. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  10. The yeast DNA repair gene RAD6 encodes a ubiquitin-conjugating enzyme.
    Nature. 1987 Sep 10-16;329(6135):131-4 PMID: 3306404
  11. Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae.
    Microbiol Rev. 1988 Mar;52(1):70-102 PMID: 3280967
  12. Cloning and expression of the OMP decarboxylase gene URA4 from Schizosaccharomyces pombe.
    Curr Genet. 1987;12(7):527-34 PMID: 2834100
  13. Domain structure and functional analysis of the carboxyl-terminal polyacidic sequence of the RAD6 protein of Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Mar;8(3):1179-85 PMID: 3285176
  14. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins.
    Science. 1988 Jun 24;240(4860):1759-64 PMID: 3289117
  15. The yeast cell cycle gene CDC34 encodes a ubiquitin-conjugating enzyme.
    Science. 1988 Sep 9;241(4871):1331-5 PMID: 2842867
  16. How eukaryotic transcriptional activators work.
    Nature. 1988 Oct 20;335(6192):683-9 PMID: 3050531
  17. A yeast DNA repair gene partially complements defective excision repair in mammalian cells.
    EMBO J. 1988 Oct;7(10):3245-53 PMID: 3053162
  18. The scanning model for translation: an update.
    J Cell Biol. 1989 Feb;108(2):229-41 PMID: 2645293
  19. Construction of a Not I restriction map of the fission yeast Schizosaccharomyces pombe genome.
    Nucleic Acids Res. 1989 Apr 11;17(7):2801-18 PMID: 2785683
  20. The Saccharomyces cerevisiae RAD2 gene complements a Schizosaccharomyces pombe repair mutation.
    Curr Genet. 1989 Jan;15(1):27-30 PMID: 2663184
  21. Isolation and initial characterization of a Schizosaccharomyces pombe mutant exhibiting temperature-dependent radiation sensitivity due to a mutation in a previously unidentified rad locus.
    Mol Gen Genet. 1989 Sep;218(3):554-8 PMID: 2586486
  22. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  23. Contribution of a caffeine-sensitive recombinational repair pathway to survival and mutagenesis in UV-irradiated Schizosaccharomyces pombe.
    Mol Gen Genet. 1978 Nov 16;167(1):43-9 PMID: 739977
  24. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene.
    Gene. 1979 Dec;8(1):121-33 PMID: 395029
  25. Both caffeine-induced lethality and the negative liquid holding effect, in UV- or gamma-irradiated wild-type Schizosaccharomyces pombe, are consequences of interference with a recombinational repair process.
    Mol Gen Genet. 1981;181(3):283-7 PMID: 6941064
  26. Spontaneous and UV-induced recombination in radiation-sensitive mutants of Schizosaccharomyces pombe.
    Mutat Res. 1981 Mar;81(1):37-48 PMID: 7254221
  27. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.
    J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
  28. Construction of a Schizosaccharomyces pombe gene bank in a yeast bacterial shuttle vector and its use to isolate genes by complementation.
    Mol Gen Genet. 1982;187(2):326-9 PMID: 6294466
  29. Plasmids carrying the yeast OMP decarboxylase structural and regulatory genes: transcription regulation in a foreign environment.
    Cell. 1983 Feb;32(2):371-7 PMID: 6337726
  30. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  31. Graphic methods to determine the function of nucleic acid sequences.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):521-38 PMID: 6364040
  32. RAD6 gene of Saccharomyces cerevisiae encodes a protein containing a tract of 13 consecutive aspartates.
    Proc Natl Acad Sci U S A. 1985 Jan;82(1):168-72 PMID: 3881753
  33. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92 PMID: 3881765
  34. Primary structure of the RAD52 gene in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Dec;4(12):2735-44 PMID: 6098821
  35. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074-8 PMID: 3156376
  36. RAD3 gene of Saccharomyces cerevisiae: nucleotide sequence of wild-type and mutant alleles, transcript mapping, and aspects of gene regulation.
    Mol Cell Biol. 1985 Jan;5(1):17-26 PMID: 3885009
  37. Recovery, repair, and mutagenesis in Schizosaccharomyces pombe.
    Adv Genet. 1985;23:1-72 PMID: 3887858
  38. Molecular cloning and characterization of the yeast RAD10 gene and expression of RAD10 protein in E. coli.
    EMBO J. 1985 Jun;4(6):1575-82 PMID: 3896774
  39. The RAD2 gene of Saccharomyces cerevisiae: nucleotide sequence and transcript mapping.
    Gene. 1985;36(3):225-34 PMID: 3000874
  40. Isolation and characterization of the structural gene for secreted acid phosphatase from Schizosaccharomyces pombe.
    J Biol Chem. 1986 Feb 25;261(6):2936-41 PMID: 3005272
  41. Analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8253-7 PMID: 3464952
  42. Nucleotide sequence and functional analysis of the RAD1 gene of Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Mar;7(3):1012-20 PMID: 3550428
  43. Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2.
    Nature. 1987 May 7-13;327(6117):31-5 PMID: 3553962
  44. On the effect of caffeine on mutation and recombination in Schizosaccharomyces pombe.
    Mol Gen Genet. 1971;110(4):348-54 PMID: 5574427
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.7.3750-3760.1990
Published
1990-07-00
Pages
3750-60
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360827
Subset
IM
Grants
NCI NIH HHS · CA01062 · United States
NIGMS NIH HHS · GM31253 · United States
Databases
GENBANK
M38132
External Links
PubMed source DOI: 10.1128/mcb.10.7.3750-3760.1990 PMC Full Text View journal details
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