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The XPV (xeroderma pigmentosum variant) gene encodes human DNA polymerase η

Abstract

Xeroderma pigmentosum variant (XP-V) is an inherited disorder which is associated with increased incidence of sunlight-induced skin cancers. Unlike other xeroderma pigmentosum cells (belonging to groups XP-A to XP-G), XP-V cells carry out normal nucleotide-excision repair processes but are defective in their replication of ultraviolet-damaged DNA1,2. It has been suspected for some time that the XPV gene encodes a protein that is involved in trans-lesion DNA synthesis, but the gene product has never been isolated. Using an improved cell-free assay for trans-lesion DNA synthesis, we have recently isolated a DNA polymerase from HeLa cells that continues replication on damaged DNA by bypassing ultraviolet-induced thymine dimers in XP-V cell extracts3. Here we show that this polymerase is a human homologue of the yeast Rad30 protein, recently identified as DNA polymerase η (ref. 4). This polymerase and yeast Rad30 are members of a family of damage-bypass replication proteins5,6,7,8,9,10 which comprises the Escherichia coli proteins UmuC and DinB and the yeast Rev1 protein. We found that all XP-V cells examined carry mutations in their DNA polymerase η gene. Recombinant human DNA polymerase η corrects the inability of XP-V cell extracts to carry out DNA replication by bypassing thymine dimers on damaged DNA. Together, these results indicate that DNA polymerase η could be the XPV gene product.

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Figure 1: Deduced amino-acid sequence of XPV protein.
Figure 2: Northern-blot analysis of XPV .
Figure 3: Mutations in the XPV cDNA of XP-V cells.
Figure 4: Activity of recombinant XPV protein.

References

  1. Lehmann, A. R. et al. . Xeroderma pigmentosum cells with normal levels of excision repair have a defect in DNA synthesis after UV-irradiation. Proc. Natl Acad. Sci. USA 72, 219–23 (1975).

    Article  ADS  CAS  Google Scholar 

  2. Friedberg, E. C., Walker, G. C. & Siede, W. DNA Repair and Mutagenesis(ASM, Washington DC, 1995).

    Google Scholar 

  3. Masutani, C. et al. . Xeroderma pigmentosum variant (XP-V) correcting protein from HeLa cells has a thymine dimer bypass DNA polymerase activity. EMBO J.(in the press).

  4. Johnson, R. E., Prakash, S. & Prakash, L. Efficient bypass of a thymine–thymine dimer by yeast DNA polymerase, polη. Science 283, 1001–1004 (1999).

    Article  ADS  CAS  Google Scholar 

  5. Kim, S. R. et al. . Multiple pathways for SOS-induced mutagenesis in Escherichia coli : an overexpression of dinB/dinP results in strongly enhancing mutagenesis in the absence of any exogenous treatment to damage DNA. Proc. Natl Acad. Sci. USA 94, 13792–13797 (1997).

    Article  ADS  CAS  Google Scholar 

  6. Smith, B. T. & Walker, G. C. Mutagenesis and more: umuDC and the Escherichia coli SOS response. Genetics 148, 1599–1610 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Nelson, J. R., Lawrence, C. W. & Hinkle, D. C. Deoxycytidyl transferase activity of yeast REV1 protein. Nature 382, 729–731 (1996).

    Article  ADS  CAS  Google Scholar 

  8. McDonald, J. P., Levine, A. S. & Woodgate, R. The Saccharomyces cerevisiae RAD30 gene, a homologue of Escherichia coli dinB and umuC, is DNA damage inducible and functions in a novel error-free postreplication repair mechanism. Genetics 147, 1557–1568 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Roush, A. A. et al. . Deletion of the Saccharomyces cerevisiae gene RAD30 encoding an Escherichia coli DinB homolog confers UV radiation sensitivity and altered mutability. Mol. Gen. Genet. 257, 686–692 (1998).

    Article  CAS  Google Scholar 

  10. Walker, G. C. SOS-regulated proteins in translesion DNA synthesis and mutagenesis. Trends Biochem. Sci. 20, 416–420 (1995).

    Article  CAS  Google Scholar 

  11. Jaspers, N. G. J., Jansen-van de Kuilen, G. & Bootsma, D. Complementation analysis of xeroderma pigmentosum variants. Exp. Cell. Res. 136, 81–90 (1981).

    Article  CAS  Google Scholar 

  12. Svoboda, D. L., Briley, L. P. & Vos, J.-M. H. Defective bypass replication of a leading strand cyclobutane thymine dimer in xeroderma pigmentosum variant cell extracts. Cancer Res. 58, 2445–2448 (1998).

    CAS  PubMed  Google Scholar 

  13. Cordeiro-Stone, M., Zaritskaya, L. S., Price, L. K. & Kaufmann, W. K. Replication fork bypass of a pyrimidine dimer blocking leading strand DNA synthesis. J. Biol. Chem. 272, 13945–13954 (1997).

    Article  CAS  Google Scholar 

  14. Cordonnier, A. M., Lehmann, A. R. & Fuchs, R. P. Impaired translesion synthesis in xeroderma pigmentosum variant extracts. Mol. Cell. Biol. 19, 2206–2211 (1999).

    Article  CAS  Google Scholar 

  15. Nelson, J. R., Lawrence, C. W. & Hinkle, D. C. Thymine–thymine dimer bypass by yeast DNA polymerase ζ. Science 272, 1646–1649 (1996).

    Article  ADS  CAS  Google Scholar 

  16. Baynton, K., Bresson-Roy, A. & Fuchs, R. P. Analysis of damage tolerance pathways in Saccharomyces cerevisiae : a requirement for Rev3 DNA polymerase in translesion synthesis. Mol. Cell. Biol. 18, 960–966 (1998).

    Article  CAS  Google Scholar 

  17. Xiao, W. et al. . Identification, chromosomal mapping and tissue-specific expression of hREV3 encoding a putative human DNA polymerase ζ. Carcinogenesis 19, 945–949 (1998).

    Article  CAS  Google Scholar 

  18. Gibbs, P. E. et al. . Ahuman homolog of the Saccharomyces cerevisiae REV3 gene, which encodes the catalytic subunit of DNA polymerase ζ. Proc. Natl Acad. Sci. USA 95, 6876–6880 (1998).

    Article  ADS  CAS  Google Scholar 

  19. Wang, Y. C., Maher, V. M. & McCormick, J. J. Xeroderma pigmentosum variant cells are less likely than normal cells to incorporate dAMP opposite photoproducts during replication of UV-irradiated plasmids. Proc. Natl Acad. Sci. USA 88, 7810–7814 (1991).

    Article  ADS  CAS  Google Scholar 

  20. Wang, Y.-C., Maher, V. M., Mitchell, D. L. & McCormick, J. J. Evidence from mutation spectra that the UV hypermutability of xeroderma pigmentosum variant cells reflects abnormal, error-prone replication on a template containing photoproducts. Mol. Cell. Biol. 13, 4276–7283 (1993).

    Article  CAS  Google Scholar 

  21. McGregor, W. G., Wei, D., Maher, V. M. & McCormick, J. J. Abnormal, error-prone bypass of photoproducts by xeroderma pigmentosum variant cell extracts results in extreme strand bias for the kinds of mutations induced by UV light. Mol. Cell. Biol. 19, 147–154 (1999).

    Article  CAS  Google Scholar 

  22. Masaki, T., Tanabe, M., Nakamura, K. & Soejima, M. Studies on a new proteolytic enzyme from Achromobacter lyticus M497-1. I. Purification and some enzymatic properties. Biochim. Biophys. Acta 660, 44–50 (1981).

    Article  CAS  Google Scholar 

  23. Masutani, C. et al. . Purification and cloning of a nucleotide excision repair complex involving the xeroderma pigmentosum group C protein and human homolog of yeast RAD23. EMBO J. 13, 1831–1843 (1994).

    Article  CAS  Google Scholar 

  24. Murata, T., Iwai, S. & Ohtsuka, E. Synthesis and characterization of a substrate for T4 endonuclease V containing a phosphorodithioate linkage at the thymine dimer site. Nucleic Acids Res. 18, 7279–7286 (1990).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank members of F.H.'s laboratory for helpful discussions, K. Tanaka for XP2SA cells, and J. H. J. Hoeijmakers for XP7TA, XP1RO, XP30RO and XP4BE cells. This work was supported by grants from the Ministry of Education, Science, Sports and Culture of Japan, and from the Biodesign Research Program of the Institute of Physical and Chemical Research (RIKEN), M.Y. is a special postdoctoral researcher of RIKEN.

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Correspondence to Fumio Hanaoka.

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Masutani, C., Kusumoto, R., Yamada, A. et al. The XPV (xeroderma pigmentosum variant) gene encodes human DNA polymerase η. Nature 399, 700–704 (1999). https://doi.org/10.1038/21447

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