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. 1991 Dec 25;19(24):6737–6741. doi: 10.1093/nar/19.24.6737

Cloning and characterization of the rad4 gene of Schizosaccharomyces pombe; a gene showing short regions of sequence similarity to the human XRCC1 gene.

M Fenech 1, A M Carr 1, J Murray 1, F Z Watts 1, A R Lehmann 1
PMCID: PMC329303  PMID: 1762905

Abstract

The rad4.116 mutant of the fission yeast Schizosaccharomyces pombe is temperature-sensitive for growth, as well as being sensitive to the killing actions of both ultraviolet light and ionizing radiation. We have cloned the rad4 gene by complementation of the temperature sensitive phenotype of the rad4.116 mutant with a S. pombe gene bank. The rad4 gene fully complemented the UV sensitivity of the rad4.116 mutant. The gene is predicted to encode a protein of 579 amino acids with a basic tail, a possible zinc finger and a nuclear location signal. The amino terminal part of the predicted rad4 ORF contains two short regions of similarity to the C-terminal part of the human XRCC1 gene. Codon usage suggests that the gene is very poorly expressed, and this was confirmed by RNA studies. Gene disruption showed that the rad4 gene was essential for the mitotic growth of S. pombe.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aves S. J., Durkacz B. W., Carr A., Nurse P. Cloning, sequencing and transcriptional control of the Schizosaccharomyces pombe cdc10 'start' gene. EMBO J. 1985 Feb;4(2):457–463. doi: 10.1002/j.1460-2075.1985.tb03651.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beach D., Piper M., Nurse P. 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):326–329. doi: 10.1007/BF00331138. [DOI] [PubMed] [Google Scholar]
  3. Broughton B. C., Barbet N., Murray J., Watts F. Z., Koken M. H., Lehmann A. R., Carr A. M. Assignment of ten DNA repair genes from Schizosaccharomyces pombe to chromosomal NotI restriction fragments. Mol Gen Genet. 1991 Sep;228(3):470–472. doi: 10.1007/BF00260641. [DOI] [PubMed] [Google Scholar]
  4. Carr A. M., MacNeill S. A., Hayles J., Nurse P. Molecular cloning and sequence analysis of mutant alleles of the fission yeast cdc2 protein kinase gene: implications for cdc2+ protein structure and function. Mol Gen Genet. 1989 Jul;218(1):41–49. doi: 10.1007/BF00330563. [DOI] [PubMed] [Google Scholar]
  5. Dorado G., Steingrimsdottir H., Arlett C. F., Lehmann A. R. Molecular analysis of ultraviolet-induced mutations in a xeroderma pigmentosum cell line. J Mol Biol. 1991 Jan 20;217(2):217–222. doi: 10.1016/0022-2836(91)90533-c. [DOI] [PubMed] [Google Scholar]
  6. Duck P., Nasim A., James A. P. Temperature-sensitive mutant of Schizosaccharomyces pombe exhibiting enhanced radiation sensitivity. J Bacteriol. 1976 Nov;128(2):536–539. doi: 10.1128/jb.128.2.536-539.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Durkacz B., Carr A., Nurse P. Transcription of the cdc2 cell cycle control gene of the fission yeast Schizosaccharomyces pombe. EMBO J. 1986 Feb;5(2):369–373. doi: 10.1002/j.1460-2075.1986.tb04221.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fan J. B., Chikashige Y., Smith C. L., Niwa O., Yanagida M., Cantor C. R. Construction of a Not I restriction map of the fission yeast Schizosaccharomyces pombe genome. Nucleic Acids Res. 1989 Apr 11;17(7):2801–2818. doi: 10.1093/nar/17.7.2801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedberg E. C. Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1988 Mar;52(1):70–102. doi: 10.1128/mr.52.1.70-102.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gentner N. E., Werner M. M., Hannan M. A., Nasim A. 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):43–49. doi: 10.1007/BF00270320. [DOI] [PubMed] [Google Scholar]
  11. Grimm C., Kohli J., Murray J., Maundrell K. Genetic engineering of Schizosaccharomyces pombe: a system for gene disruption and replacement using the ura4 gene as a selectable marker. Mol Gen Genet. 1988 Dec;215(1):81–86. doi: 10.1007/BF00331307. [DOI] [PubMed] [Google Scholar]
  12. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  13. Lieberman H. B., Riley R., Martel M. 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):554–558. doi: 10.1007/BF00332423. [DOI] [PubMed] [Google Scholar]
  14. Murray J. M., Carr A. M., Lehmann A. R., Watts F. Z. Cloning and characterisation of the rad9 DNA repair gene from Schizosaccharomyces pombe. Nucleic Acids Res. 1991 Jul 11;19(13):3525–3531. doi: 10.1093/nar/19.13.3525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nurse P. Genetic control of cell size at cell division in yeast. Nature. 1975 Aug 14;256(5518):547–551. doi: 10.1038/256547a0. [DOI] [PubMed] [Google Scholar]
  16. Phipps J., Nasim A., Miller D. R. Recovery, repair, and mutagenesis in Schizosaccharomyces pombe. Adv Genet. 1985;23:1–72. doi: 10.1016/s0065-2660(08)60511-8. [DOI] [PubMed] [Google Scholar]
  17. Sunnerhagen P., Seaton B. L., Nasim A., Subramani S. Cloning and analysis of a gene involved in DNA repair and recombination, the rad1 gene of Schizosaccharomyces pombe. Mol Cell Biol. 1990 Jul;10(7):3750–3760. doi: 10.1128/mcb.10.7.3750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thompson L. H., Brookman K. W., Dillehay L. E., Carrano A. V., Mazrimas J. A., Mooney C. L., Minkler J. L. A CHO-cell strain having hypersensitivity to mutagens, a defect in DNA strand-break repair, and an extraordinary baseline frequency of sister-chromatid exchange. Mutat Res. 1982 Aug;95(2-3):427–440. doi: 10.1016/0027-5107(82)90276-7. [DOI] [PubMed] [Google Scholar]
  19. Thompson L. H., Brookman K. W., Jones N. J., Allen S. A., Carrano A. V. Molecular cloning of the human XRCC1 gene, which corrects defective DNA strand break repair and sister chromatid exchange. Mol Cell Biol. 1990 Dec;10(12):6160–6171. doi: 10.1128/mcb.10.12.6160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wright A., Maundrell K., Heyer W. D., Beach D., Nurse P. Vectors for the construction of gene banks and the integration of cloned genes in Schizosaccharomyces pombe and Saccharomyces cerevisiae. Plasmid. 1986 Mar;15(2):156–158. doi: 10.1016/0147-619x(86)90051-x. [DOI] [PubMed] [Google Scholar]

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