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. 1990 Sep;87(17):6679–6683. doi: 10.1073/pnas.87.17.6679

Human DNA ligase I cDNA: cloning and functional expression in Saccharomyces cerevisiae.

D E Barnes 1, L H Johnston 1, K Kodama 1, A E Tomkinson 1, D D Lasko 1, T Lindahl 1
PMCID: PMC54600  PMID: 2204063

Abstract

Human cDNA clones encoding the major DNA ligase activity in proliferating cells, DNA ligase I, were isolated by two independent methods. In one approach, a human cDNA library was screened by hybridization with oligonucleotides deduced from partial amino acid sequence of purified bovine DNA ligase I. In an alternative approach, a human cDNA library was screened for functional expression of a polypeptide able to complement a cdc9 temperature-sensitive DNA ligase mutant of Saccharomyces cerevisiae. The sequence of an apparently full-length cDNA encodes a 102-kDa protein, indistinguishable in size from authentic human DNA ligase I. The deduced amino acid sequence of the human DNA ligase I cDNA is 40% homologous to the smaller DNA ligases of S. cerevisiae and Schizosaccharomyces pombe, homology being confined to the carboxyl-terminal regions of the respective proteins. Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is transcribed from a single-copy gene on chromosome 19.

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Selected References

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

  1. Arrand J. E., Willis A. E., Goldsmith I., Lindahl T. Different substrate specificities of the two DNA ligases of mammalian cells. J Biol Chem. 1986 Jul 15;261(20):9079–9082. [PubMed] [Google Scholar]
  2. Barker D. G., Johnson A. L., Johnston L. H. An improved assay for DNA ligase reveals temperature-sensitive activity in cdc9 mutants of Saccharomyces cerevisiae. Mol Gen Genet. 1985;200(3):458–462. doi: 10.1007/BF00425731. [DOI] [PubMed] [Google Scholar]
  3. Barker D. G., Johnston L. H. Saccharomyces cerevisiae cdc9, a structural gene for yeast DNA ligase which complements Schizosaccharomyces pombe cdc17. Eur J Biochem. 1983 Aug 1;134(2):315–319. doi: 10.1111/j.1432-1033.1983.tb07568.x. [DOI] [PubMed] [Google Scholar]
  4. Barker D. G., White J. H., Johnston L. H. Molecular characterisation of the DNA ligase gene, CDC17, from the fission yeast Schizosaccharomyces pombe. Eur J Biochem. 1987 Feb 2;162(3):659–667. doi: 10.1111/j.1432-1033.1987.tb10688.x. [DOI] [PubMed] [Google Scholar]
  5. D'Arpa P., Machlin P. S., Ratrie H., 3rd, Rothfield N. F., Cleveland D. W., Earnshaw W. C. cDNA cloning of human DNA topoisomerase I: catalytic activity of a 67.7-kDa carboxyl-terminal fragment. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2543–2547. doi: 10.1073/pnas.85.8.2543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edwards Y. H., Parkar M., Povey S., West L. F., Parrington J. M., Solomon E. Human myosin heavy chain genes assigned to chromosome 17 using a human cDNA clone as probe. Ann Hum Genet. 1985 May;49(Pt 2):101–109. doi: 10.1111/j.1469-1809.1985.tb01681.x. [DOI] [PubMed] [Google Scholar]
  7. Fabre F., Roman H. Evidence that a single DNA ligase is involved in replication and recombination in yeast. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4586–4588. doi: 10.1073/pnas.76.9.4586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heisterkamp N., Groffen J., Stephenson J. R., Spurr N. K., Goodfellow P. N., Solomon E., Carritt B., Bodmer W. F. Chromosomal localization of human cellular homologues of two viral oncogenes. Nature. 1982 Oct 21;299(5885):747–749. doi: 10.1038/299747a0. [DOI] [PubMed] [Google Scholar]
  9. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnston L. H., Barker D. G., Nurse P. Cloning and characterization of the Schizosaccharomyces pombe DNA ligase gene CDC17. Gene. 1986;41(2-3):321–325. doi: 10.1016/0378-1119(86)90114-9. [DOI] [PubMed] [Google Scholar]
  11. Johnston L. H., Nasmyth K. A. Saccharomyces cerevisiae cell cycle mutant cdc9 is defective in DNA ligase. Nature. 1978 Aug 31;274(5674):891–893. doi: 10.1038/274891a0. [DOI] [PubMed] [Google Scholar]
  12. Johnston L. H. The DNA repair capability of cdc9, the Saccharomyces cerevisiae mutant defective in DNA ligase. Mol Gen Genet. 1979 Feb 16;170(1):89–92. doi: 10.1007/BF00268583. [DOI] [PubMed] [Google Scholar]
  13. Kielty C. M., Povey S., Hopkinson D. A. Regulation of expression of liver-specific enzymes. III. Further analysis of a series of rat hepatoma X human somatic cell hybrids. Ann Hum Genet. 1982 Oct;46(Pt 4):307–327. doi: 10.1111/j.1469-1809.1982.tb01582.x. [DOI] [PubMed] [Google Scholar]
  14. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lasko D. D., Tomkinson A. E., Lindahl T. Mammalian DNA ligases. Biosynthesis and intracellular localization of DNA ligase I. J Biol Chem. 1990 Jul 25;265(21):12618–12622. [PubMed] [Google Scholar]
  16. Lathe R. Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations. J Mol Biol. 1985 May 5;183(1):1–12. doi: 10.1016/0022-2836(85)90276-1. [DOI] [PubMed] [Google Scholar]
  17. Lee M. G., Nurse P. Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2. Nature. 1987 May 7;327(6117):31–35. doi: 10.1038/327031a0. [DOI] [PubMed] [Google Scholar]
  18. Nabholz M., Miggiano V., Bodmer W. Genetic analysis with human--mouse somatic cell hybrids. Nature. 1969 Jul 26;223(5204):358–363. doi: 10.1038/223358a0. [DOI] [PubMed] [Google Scholar]
  19. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  20. Olsen L. C., Aasland R., Wittwer C. U., Krokan H. E., Helland D. E. Molecular cloning of human uracil-DNA glycosylase, a highly conserved DNA repair enzyme. EMBO J. 1989 Oct;8(10):3121–3125. doi: 10.1002/j.1460-2075.1989.tb08464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Phillips I. R., Shephard E. A., Povey S., Davis M. B., Kelsey G., Monteiro M., West L. F., Cowell J. A cytochrome P-450 gene family mapped to human chromosome 19. Ann Hum Genet. 1985 Oct;49(Pt 4):267–274. doi: 10.1111/j.1469-1809.1985.tb01702.x. [DOI] [PubMed] [Google Scholar]
  22. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Silver P., Goodson H. Nuclear protein transport. Crit Rev Biochem Mol Biol. 1989;24(4):419–435. doi: 10.3109/10409238909082557. [DOI] [PubMed] [Google Scholar]
  25. Smith G. L., Chan Y. S., Kerr S. M. Transcriptional mapping and nucleotide sequence of a vaccinia virus gene encoding a polypeptide with extensive homology to DNA ligases. Nucleic Acids Res. 1989 Nov 25;17(22):9051–9062. doi: 10.1093/nar/17.22.9051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Solomon E., Bobrow M., Goodfellow P. N., Bodmer W. F., Swallow D. M., Povey S., Noël B. Human gene mapping using an X/autosome translocation. Somatic Cell Genet. 1976 Mar;2(2):125–140. doi: 10.1007/BF01542626. [DOI] [PubMed] [Google Scholar]
  27. Solomon E., Swallow D., Burgess S., Evans L. Assignment of the human acid alpha-glucosidase gene (alphaGLU) to chromosome 17 using somatic cell hybrids. Ann Hum Genet. 1979 Jan;42(3):273–281. doi: 10.1111/j.1469-1809.1979.tb00661.x. [DOI] [PubMed] [Google Scholar]
  28. Swallow D. M., Solomon E., Pajunen L. Immunochemical analysis of the N-acetyl hexosaminidases in human-mouse hybrids made using a double selective system. Cytogenet Cell Genet. 1977;18(3):136–148. doi: 10.1159/000130758. [DOI] [PubMed] [Google Scholar]
  29. Söderhäll S., Lindahl T. Mammalian DNA ligases. Serological evidence for two separate enzymes. J Biol Chem. 1975 Nov 10;250(21):8438–8444. [PubMed] [Google Scholar]
  30. Teraoka H., Tsukada K. Immunochemical analysis of molecular forms of mammalian DNA ligases I and II. Biochim Biophys Acta. 1986 Sep 26;873(2):297–303. doi: 10.1016/0167-4838(86)90057-9. [DOI] [PubMed] [Google Scholar]
  31. Tomkinson A. E., Lasko D. D., Daly G., Lindahl T. Mammalian DNA ligases. Catalytic domain and size of DNA ligase I. J Biol Chem. 1990 Jul 25;265(21):12611–12617. [PubMed] [Google Scholar]
  32. Whitehead A. S., Solomon E., Chambers S., Bodmer W. F., Povey S., Fey G. Assignment of the structural gene for the third component of human complement to chromosome 19. Proc Natl Acad Sci U S A. 1982 Aug;79(16):5021–5025. doi: 10.1073/pnas.79.16.5021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Willis A. E., Weksberg R., Tomlinson S., Lindahl T. Structural alterations of DNA ligase I in Bloom syndrome. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8016–8020. doi: 10.1073/pnas.84.22.8016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wong S. W., Wahl A. F., Yuan P. M., Arai N., Pearson B. E., Arai K., Korn D., Hunkapiller M. W., Wang T. S. Human DNA polymerase alpha gene expression is cell proliferation dependent and its primary structure is similar to both prokaryotic and eukaryotic replicative DNA polymerases. EMBO J. 1988 Jan;7(1):37–47. doi: 10.1002/j.1460-2075.1988.tb02781.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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