Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Characterization of human exonuclease 1 in complex with mismatch repair proteins, subcellular localization and association with PCNA

Oncogene volume 23, pages 1457–1468 (2004)Cite this article

Abstract

Human exonuclease 1 (hEXO1) has been implicated in DNA mismatch repair (MMR), replication, and recombination, but the nature of its interaction with these cellular processes is still ambiguous. We show that hEXO1 colocalizes with proliferating cell nuclear antigen (PCNA) at DNA replication sites and that the C-terminal region of hEXO1 is sufficient for this localization. We also show that both hMLH1–hPMS2 (MutLα) and hMLH1–hEXO1 complexes are formed in a reaction mixture containing all three proteins. Moreover, hEXO1 5′ double-stranded exonuclease activity on a homoduplex substrate but not on a substrate containing a G/T mismatch was inhibited by complex formation with hMSH2–hMSH6 (MutSα) or MutLα. Taken together, the results support a model in which hEXO1 plays a role in events at the replication sites as well as a functional role in the MMR and/or recombination processes.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  • Amin NS, Nguyen M-N, Oh S and Kolodner RD . (2001). Mol. Cell. Biol., 21, 5142–5155.

  • Aquilina G and Bignami M . (2001). J. Cell. Physiol., 187, 145–154.

    Article  CAS  Google Scholar 

  • Argueso JL, Smith D, Yi J, Waase M, Sarin S and Alani E . (2002). Genetics, 160, 909–921.

  • Baker SM, Bronner CE, Zhang L, Plug AW, Robatzek M, Warren G, Elliott EA, Yu J, Ashley T, Arnheim N, Flavell RA and Liskay RM . (1995). Cell, 82, 309–319.

    Article  CAS  Google Scholar 

  • Baker SM, Harris AC, Tsao JL, Flath TJ, Bronner CE, Gordon M, Shibata D and Liskay RM . (1998). Cancer Res., 58, 1087–1089.

  • Bravo R, Frank R, Blundell PA and Macdonald-Bravo H . (1987). Nature, 326, 515–517.

    Article  CAS  Google Scholar 

  • Bravo R and Macdonald-Bravo H . (1987). J. Cell. Biol., 105, 1549–1554.

  • Brénot-Bosc F, Gupta S, Margolis RL and Fotedar R . (1995). Chromosoma, 103, 517–527.

  • Buermeyer AB, Deschênes SM, Baker SM and Liskay RM . (1999). Annu. Rev. Genet., 33, 533–564.

    Article  CAS  Google Scholar 

  • Cao L, Alani E and Kleckner N . (1990). Cell, 61, 1089–1101.

    Article  CAS  Google Scholar 

  • Celis JE and Celis A . (1985). Proc. Natl. Acad. Sci. USA, 82, 3262–3266.

  • Datta A, Schmeits JL, Amin NS, Lau PJ, Myung K and Kolodner RD . (2000). Mol. Cell., 6, 593–603.

  • Digilio FA, Pannuti A, Lucchesi JC, Furia M and Polito LC . (1996). Dev. Biol., 178, 90–100.

  • Drotschmann K, Clark AB, Tran HT, Resnick MA, Gordenin DA and Kunkel TA . (1999). Proc. Natl. Acad. Sci. USA, 96, 2970–2975.

  • Edelmann W, Cohen PE, Kane M, Lau K, Morrow B, Bennett S, Umar A, Kunkel T, Cattoretti G, Chaganti R, Pollard JW, Kolodner RD and Kucherlapati R . (1996). Cell, 85, 1125–1134.

    Article  CAS  Google Scholar 

  • Edelmann W, Umar A, Yang K, Heyer J, Kucherlapati M, Lia M, Kneitz B, Avdievich E, Fan K, Wong E, Crouse G, Kunkel T, Lipkin M, Kolodner RD and Kucherlapati R . (2000). Cancer Res., 60, 803–807.

  • Edelmann W, Yang K, Umar A, Heyer J, Lau K, Fan K, Liedtke W, Cohen PE, Kane MF, Lipford JR, Yu N, Crouse GF, Pollard JW, Kunkel T, Lipkin M, Kolodner R and Kucherlapati R . (1997). Cell, 91, 467–477.

    Article  CAS  Google Scholar 

  • Fiorentini P, Huang KN, Tishkoff DX, Kolodner RD and Symington LS . (1997). Mol. Cell. Biol., 17, 2764–2773.

    Article  CAS  Google Scholar 

  • Fuss J and Linn S . (2002). J. Biol. Chem., 277, 8658–8666.

  • Genschel J, Bazemore LR and Modrich P . (2002). J. Biol. Chem., 277, 13302–13311.

    Article  CAS  Google Scholar 

  • Heyer J, Yang K, Lipkin M, Edelmann W and Kucherlapati R . (1999). Oncogene, 18, 5325–5333.

    Article  CAS  Google Scholar 

  • Hozak P, Hassan AB, Jackson DA and Cook PR . (1993). Cell, 73, 361–373.

    Article  Google Scholar 

  • Hsieh P . (2001). Mutat. Res., 486, 71–87.

  • Jäger AC, Rasmussen M, Bisgaard HC, Singh KK, Nielsen FC and Rasmussen LJ . (2001). Oncogene, 20, 3590–3595.

    Article  Google Scholar 

  • Kamath-Loeb AS, Loeb LA, Johansson E, Burgers PM and Fry M . (2001). J. Biol. Chem., 276, 16439–16446.

    Article  CAS  Google Scholar 

  • Karran P . (2001). Carcinogenesis, 22, 1931–1937.

    Article  CAS  Google Scholar 

  • Kawamura K, Kobayashi Y, Tanaka T, Ikeda R, Fujikawa-Yamamoto K and Suzuki K . (2000). Anal. Quant. Cytol. Histol., 22, 107–113.

  • Khazanehdari KA and Borts RH . (2000). Chromosoma, 109, 94–102.

    Article  CAS  Google Scholar 

  • Kirkpatrick DT, Ferguson JR, Petes TD and Symington LS . (2000). Genetics, 156, 1549–1557.

  • Kleczkowska HE, Marra G, Lettieri T and Jiricny J . (2001). Genes Dev., 15, 724–736.

  • Kondo E, Horii A and Fukushige S . (2001). Nucleic Acids Res., 29, 1695–1702.

  • Leach FS, Polyak K, Burrell M, Johnson KA, Hill D, Dunlop MG, Wyllie AH, Peltomaki P, de la Chapelle A, Hamilton SR, Kinzler KW and Vogelstein B . (1996). Cancer Res., 56, 235–240.

  • Lebel M . (2001). Cell. Mol. Life Sci., 58, 857–867.

  • Lee B-I, Shannon M, Stubbs L and Wilson DM . (1999). Nucleic Acids Res., 27, 4114–4120.

  • Lee B-I and Wilson DM . (1999). J. Biol. Chem., 274, 37763–37769.

    Article  CAS  Google Scholar 

  • Lengauer C, Kinzler KW and Vogelstein B . (1998). Nature, 396, 643–649.

    Article  CAS  Google Scholar 

  • Leonhardt H, Rahn H-P, Weinzierl P, Sporbert A, Cremer T, Zink D and Cardoso MC . (2000). J. Cell. Biol., 149, 271–279.

  • Lin YL, Shivji MK, Chen C, Kolodner R, Wood RD and Dutta A . (1998). J. Biol. Chem., 273, 1453–1461.

    Article  CAS  Google Scholar 

  • Marra G, Chang CL, Laghi LA, Chauhan DP, Young D and Boland CR . (1996). Oncogene, 13, 2189–2196.

  • Marra G and Schär P . (1999). Biochem. J., 338, 1–13.

    Article  CAS  Google Scholar 

  • Michel B, Flores MJ, Viguera E, Grompone G, Seigneur M and Bidnenko V . (2001). Proc. Natl. Acad. Sci. USA, 98, 8181–8188.

  • Modrich P and Lahue R . (1996). Annu. Rev. Biochem., 65, 101–133.

    Article  CAS  Google Scholar 

  • Nichols AF and Sancar A . (1992). Nucleic Acids Res., 20, 2441–2446.

  • Nielsen J, Christiansen J, Lykke-Andersen J, Johnsen AH, Wewer UM and Nielsen FC . (1999). Mol. Cell. Biol., 19, 1262–1270.

    Article  CAS  Google Scholar 

  • Nielsen FC, Nielsen J, Kristensen MA, Koch G and Christiansen J . (2002). J. Cell. Sci., 115, 2087–2097.

  • O'Keefe RT, Henderson SC and Spector DL . (1992). J. Cell. Biol., 116, 1095–1110.

  • Prolla TA, Baker SM, Harris AC, Tsao JL, Yao X, Bronner CE, Zheng B, Gordon M, Reneker J, Arnheim N, Shibata D, Bradley A and Liskay RM . (1998). Nat. Genet., 18, 276–279.

  • Rasmussen LJ, Rasmussen M, Lee B-I, Rasmussen AK, Wilson DM, Nielsen FC and Bisgaard HC . (2000a). Mutat. Res., 460, 41–52.

  • Rasmussen LJ, Rasmussen M, Lützen A, Bisgaard HC and Singh KK . (2000b). Exp. Cell. Res., 257, 127–134.

  • Reitmair AH, Schmits R, Ewel A, Bapat B, Redston M, Mitri A, Waterhouse P, Mittrucker HW, Wakeham A, Liu B, Thomason A, Griesser H, Gallinger S, Ballhausen WG, Fishel R and Mak TW . (1995). Nat. Genet., 11, 64–70.

  • Rothstein R, Michel B and Gangloff S . (2000). Genes Dev., 14, 1–10.

  • Schmutte C, Marinescu RC, Sadoff MM, Guerrette S, Overhauser J and Fishel R . (1998). Cancer Res., 58, 4537–4542.

  • Schmutte C, Sadoff MM, Shim KS, Acharya S and Fishel R . (2001). J. Biol. Chem., 276, 33011–33018.

    Article  CAS  Google Scholar 

  • Sharma S, Sommers JA, Driscoll HC, Uzdilla L, Wilson TM and Brosh Jr RM . (2003). J. Biol. Chem., 278, 23487–23496.

    Article  CAS  Google Scholar 

  • Shcherbakova PV, Hall MC, Lewis MS, Bennett SE, Martin KJ, Bushel PR, Afshari CA and Kunkel TA . (2001). Mol. Cell. Biol., 21, 940–951.

  • Smith BT, Grossman AD and Walker GC . (2001). Mol. Cell, 8, 1197–1206.

  • Sokolsky T and Alani E . (2000). Genetics, 155, 589–599.

  • Somanathan S, Suchyna TM, Siegel AJ and Berezney R . (2001). J. Cell. Biochem., 81, 56–67.

  • Sun H, Treco D and Szostak J . (1991). Cell, 64, 1155–1161.

    Article  CAS  Google Scholar 

  • Sun X, Zheng L and Shen B . (2002). Cancer Res., 62, 6026–6030.

  • Szankasi P and Smith GR . (1995). Science, 267, 1166–1169.

    Article  CAS  Google Scholar 

  • Thomas DC, Umar A and Kunkel TA . (1995). Methods, 7, 187–197.

    Article  CAS  Google Scholar 

  • Tishkoff DX, Amin NS, Viars CS, Arden KC and Kolodner RD . (1998). Cancer Res., 58, 5027–5031.

  • Tishkoff DX, Boerger AL, Bertrand P, Filosi N, Gaida GM, Kane MF and Kolodner RD . (1997). Proc. Natl. Acad. Sci. USA, 94, 7487–7492.

  • Tran HT, Gordenin DA and Resnick MA . (1999). Mol. Cell. Biol., 19, 2000–2007.

    Article  CAS  Google Scholar 

  • Tran PT, Erdeniz N, Dudley S and Liskay RM . (2002). DNA Repair, 1, 895–912.

    Article  CAS  Google Scholar 

  • Tsubouchi H and Ogawa H . (2000). Mol. Biol. Cell, 11, 2221–2233.

  • Wang Y, Cortez D, Yazdi P, Neff N, Elledge SJ and Qin J . (2000). Genes Dev., 14, 927–939.

  • Wilson DM, Carney JP, Coleman MA, Adamson AW, Christensen M and Lamerdin JE . (1998). Nucleic Acids Res., 26, 3762–3768.

  • de Wind N, Dekker M, Berns A, Radman M and te Riele H . (1995). Cell, 82, 321–330.

    Article  CAS  Google Scholar 

  • de Wind N, Dekker M, Claij N, Jansen L, van Klink Y, Radman M, Riggins G, van der Valk M, van't Wout K and te Riele H . (1999). Nat. Genet., 23, 359–362.

  • de Wind N and Hays JB . (2001). Curr. Biol., 11, R545–R548.

  • Worth L, Clark S, Radman M and Modrich P . (1994). Proc. Natl. Acad. Sci. USA, 91, 3238–3241.

  • Wu Y, Berends MJW, Post JG, Mensink RGJ, Verlind E, van der Sluis T, Kempinga C, Sijmons RH, van der Zee AGJ, Hollema H, Kleibeuker JH, Buys CHCM and Hofstra RMW . (2001). Gastroenterology, 120, 1580–1587.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to Anne Marie Bundgaard, Gerda Olesen, Lene Markussen, Bente Rotbøl, and Joan Christiansen for expert technical assistance. We are grateful to Dr Jochen Genschel and Dr Poul Modrich for the generous gift of antibodies against hEXO1 and to Dr David M Wilson III and Dr Christina Cardoso for plasmids. We are also grateful to Drs Niels de Wind and Andrew Buermeyer for murine cell lines. The Danish Cancer Society (LJR), Danish Research Council (LJR), Villum Kann Rasmussen Foundation (ACJ), the NOVO-Nordisk foundation (FCN), and the John and Birthe Meier Foundation (FCN) supported this work.

Author information

Authors and Affiliations

  1. Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, DK-2100, Denmark

    Finn Cilius Nielsen, Anne Charlotte Jäger & Jens R Bundgaard

  2. Department of Life Sciences and Chemistry, Roskilde University, Roskilde, DK-4000, Denmark

    Anne Charlotte Jäger, Anne Lützen & Lene Juel Rasmussen

Authors
  1. Finn Cilius Nielsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anne Charlotte Jäger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anne Lützen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jens R Bundgaard
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lene Juel Rasmussen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lene Juel Rasmussen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nielsen, F., Jäger, A., Lützen, A. et al. Characterization of human exonuclease 1 in complex with mismatch repair proteins, subcellular localization and association with PCNA. Oncogene 23, 1457–1468 (2004). https://doi.org/10.1038/sj.onc.1207265

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1207265

Keywords

This article is cited by

Search

Advanced search

Quick links