Abstract
The DNA damage checkpoint kinase, CHK2, promotes growth arrest or apoptosis through phosphorylating targets such as Cdc25A, Cdc25C, BRCA1, and p53. Both germline and somatic loss-of-function CHEK2 mutations occur in human tumours, the former linked to the Li–Fraumeni syndrome, and the latter found in diverse types of sporadic malignancies. Here we examined the status of CHK2 by genetic and immunohistochemical analyses in 53 breast carcinomas previously characterized for TP53 status. We identified two CHEK2 mutants, 470T>C (Ile157Thr), and a novel mutation, 1368insA leading to a premature stop codon in exon 13. The truncated protein encoded by CHEK2 carrying the 1368insA was stable yet mislocalized to the cytoplasm in tumour sections and when ectopically expressed in cultured cells. Unexpectedly, we found CHEK2 to be subject to extensive alternative splicing, with some 90 splice variants detected in our tumour series. While all cancers expressed normal-length CHEK2 mRNA together with the spliced transcripts, we demonstrate and/or predict some of these splice variants to lack CHK2 function and/or localize aberrantly. We conclude that cytoplasmic sequestration may represent a novel mechanism to disable CHK2, and propose to further explore the significance of the complex splicing patterns of this tumour suppressor gene in oncogenesis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ahn JY, Li XH, Davis HL and Canman CE . (2002). J. Biol. Chem., 277, 19389–19395.
Allinen M, Huusko P, Mantyniemi S, Launonen V and Winqvist R . (2001). Br. J Cancer, 85, 209–212.
Bartek J, Falck J and Lukas J . (2001). Nat. Rev. Mol. Cell Biol., 2, 877–886.
Bartek J and Lukas J . (2003). Cancer Cell, 3, 421–429.
Bartel F, Pinkert D, Fiedler W, Kappler M, Wurl P, Schmidt H and Taubert H . (2004). Int. J. Oncol., 24, 143–151.
Bartkova J, Falck J, Rajpert-De Meyts E, Skakkebaek NE, Lukas J and Bartek J . (2001). Oncogene, 20, 5897–5902.
Bell DW, Varley JM, Szydlo TE, Kang DH, Wahrer DCR, Shannon KE, Lubratovich M, Verselis SJ, Isselbacher KJ, Fraumeni JF, Birch JM, Li FP, Garber JE and Haber DA . (1999). Science, 286, 2528–2531.
Caspari T . (2000). Curr. Biol., 10, R315–R317.
Chehab NH, Malikzay A, Appel M and Halazonetis TD . (2000). Genes Dev., 14, 278–288.
Craig A, Scott M, Burch L, Smith G, Ball K and Hupp T . (2003). EMBO Rep., 4, 787–792.
Dong X, Wang L, Taniguchi K, Wang X, Cunningham JM, McDonnell SK, Qian C, Marks AF, Slager SL, Peterson BJ, Smith DI, Cheville JC, Blute ML, Jacobsen SJ, Schaid DJ, Tindall DJ, Thibodeau SN and Liu W . (2003). Am. J. Hum. Genet., 72, 270–280.
Evdokiou A, Atkins GJ, Bouralexis S, Hay S, Raggatt LJ, Cowled PA, Graves SE, Clayer M and Findlay DM . (2001). Int. J. Oncol., 19, 625–632.
Falck J, Lukas C, Protopopova M, Lukas J, Selivanova G and Bartek J . (2001a). Oncogene, 20, 5503–5510.
Falck J, Mailand N, Syljuasen RG, Bartek J and Lukas J . (2001b). Nature, 410, 842–847.
Frebourg T, Ebel A, Bonaiti-Pellie C, Brugieres L, Berthet P, Bressac-De Paillerets B, Chevrier A, Chompret A, Cohen-Haguenauer O, Delattre O, Feingold J, Feunteun J, Frappaz D, Fricker JP, Gesta P, Jonveaux P, Kalifa C, Lasset C, Leheup B, Limacher JM, Longy M, Nogues C, Oppenheim D, Sommelet D, Soubrier F, Stoll C, Stoppa-Lyonnet D and Tristant H . (2001). Bull. Cancer, 88, 581–587.
Geisler S, Børresen-Dale A-L, Johnsen H, Aas T, Geisler J, Akslen LA, Anker G and Lønning PE . (2003). Clin. Cancer Res., 9, 5582–5588.
Geisler S, Lønning PE, Aas T, Johnsen H, Fluge O, Haugen DF, Lillehaug JR, Akslen LA and Borresen-Dale AL . (2001). Cancer Res., 61, 2505–2512.
Hirao A, Cheung A, Duncan G, Girard PM, Elia AJ, Wakeham A, Okada H, Sarkissian T, Wong JA, Sakai T, De Stanchina E, Bristow RG, Suda T, Lowe SW, Jeggo PA, Elledge SJ and Mak TW . (2002). Mol. Cell. Biol., 22, 6521–6532.
Hirao A, Kong YY, Matsuoka S, Wakeham A, Ruland J, Yoshida H, Liu D, Elledge SJ and Mak TW . (2000). Science, 287, 1824–1827.
Hofmann WK, Miller CW, Tsukasaki K, Tavor S, Ikezoe T, Hoelzer D, Takeuchi S and Koeffler HP . (2001). Leukemia Res., 25, 333–338.
Hori M, Shimazaki J, Inagawa S and Itabashi M . (2000). Pathol. Int., 50, 786–792.
Ishimoto O, Kawahara C, Enjo K, Obinata M, Nukiwa T and Ikawa S . (2002). Cancer Res., 62, 636–641.
Kraus A, Neff F, Behn M, Schuermann M, Muenkel K and Schlegel J . (1999). Int. J. Cancer, 80, 930–934.
Lee CH and Chung JH . (2001). J. Biol. Chem., 276, 30537–30541.
Lee JS, Collins KM, Brown AL, Lee CH and Chung JH . (2000). Nature, 404, 201–204.
Lee SB, Kim SH, Bell DW, Wahrer DC, Schiripo TA, Jorczak MM, Sgroi DC, Garber JE, Li FP, Nichols KE, Varley JM, Godwin AK, Shannon KM, Harlow E and Haber DA . (2001). Cancer Res., 61, 8062–8067.
Lukas C, Bartkova J, Latella L, Falck J, Mailand N, Schroeder T, Sehested M, Lukas J and Bartek J . (2001). Cancer Res., 61, 4990–4993.
Lukas C, Falck J, Bartkova J, Bartek J and Lukas J . (2003). Nat. Cell. Biol., 5, 255–260.
Lønning PE . (2003). Lancet Oncol., 4, 177–185.
Lønning PE . (2004). Trends Mol. Med., 10, 113–118.
Maier B, Gluba W, Bernier B, Turner T, Mohammad K, Guise T, Sutherland A, Thorner M and Scrable H . (2004). Genes Dev., 18, 306–319.
Matsuoka S, Huang MX and Elledge SJ . (1998). Science, 282, 1893–1897.
Matsuoka S, Nakagawa T, Masuda A, Haruki N, Elledge SJ and Takahashi T . (2001). Cancer Res., 61, 5362–5365.
Meijers-Heijboer H, van den Ouweland A, Klijn J, Wasielewski M, de Snoo A, Oldenburg R, Hollestelle A, Houben M, Crepin E, van Veghel-Plandsoen M, Elstrodt F, van Duijn C, Bartels C, Meijers C, Schutte M, McGuffog L, Thompson D, Easton D, Sodha N, Seal S, Barfoot R, Mangion J, Chang-Claude J, Eccles D, Eeles R, Evans DG, Houlston R, Murday V, Narod S, Peretz T, Peto J, Phelan C, Zhang HX, Szabo C, Devilee P, Goldgar D, Futreal PA, Nathanson KL, Weber B, Rahman N and Stratton MR . (2002). Nat. Genet., 31, 55–59.
Meijers-Heijboer H, Wijnen J, Vasen H, Wasielewski M, Wagner A, Hollestelle A, Elstrodt F, van den Bos R, de Snoo A, Fat GT, Brekelmans C, Jagmohan S, Franken P, Verkuijlen P, van den Ouweland A, Chapman P, Tops C, Moslein G, Burn J, Lynch H, Klijn J, Fodde R and Schutte M . (2003). Am. J. Hum. Genet., 72, 1308–1314.
O'Nions J, Brooks LA, Sullivan A, Bell A, Dunne B, Rozycka M, Reddy A, Tidy JA, Evans D, Farrell PJ, Evans A, Gasco M, Gusterson B and Crook T . (2001). Br. J. Cancer, 85, 1551–1556.
Reddy A, Yuille M, Sullivan A, Repellin C, Bell A, Tidy JA, Evans DJ, Farrell PJ, Gusterson B, Gasco M and Crook T . (2002). Br. J. Cancer, 86, 756–760.
Seppälä EH, Ikonen T, Mononen N, Autio V, Rokman A, Matikainen MP, Tammela TL and Schleutker J . (2003). Br. J. Cancer, 89, 1966–1970.
Shieh SY, Ahn J, Tamai K, Taya Y and Prives C . (2000). Genes Dev., 14, 289–300.
Sigalas I, Calvert AH, Anderson JJ, Neal DE and Lunec J . (1996). Nat. Med., 2, 912–917.
Sodha N, Bullock S, Taylor R, Mitchell G, Guertl-Lackner B, Williams RD, Bevan S, Bishop K, McGuire S, Houlston RS and Eeles RA . (2002a). Br. J. Cancer, 87, 1445–1448.
Sodha N, Houlston RS, Williams R, Yuille MA, Mangion J and Eeles RA . (2002b). Hum. Mutat., 19, 173–177.
Sodha N, Williams R, Mangion J, Bullock SL, Yuille MR and Eeles RA . (2000). Science, 289, 359.
Sullivan A, Yuille M, Repellin C, Reddy A, Reelfs O, Bell A, Dunne B, Gusterson BA, Osin P, Farrell PJ, Yulug I, Evans A, Ozcelik T, Gasco M and Crook T . (2002). Oncogene, 21, 1316–1324.
Takai H, Naka K, Okada Y, Watanabe M, Harada N, Saito S, Anderson CW, Appella E, Nakanishi M, Suzuki H, Nagashima K, Sawa H, Ikeda K and Motoyama N . (2002). EMBO J., 21, 5195–5205.
Theard D, Coisy M, Ducommun B, Concannon P and Darbon JM . (2001). Biochem. Biophys. Res. Commun., 289, 1199–1204.
Tominaga K, Morisaki H, Kaneko Y, Fujimoto A, Tanaka T, Ohtsubo M, Hirai M, Okayama H, Ikeda K and Nakanishi M . (1999). J. Biol. Chem., 274, 31463–31467.
Vahteristo P, Bartkova J, Eerola H, Syrjakoski K, Ojala S, Kilpivaara O, Tamminen A, Kononen J, Aittomaki K, Heikkila P, Holli K, Blomqvist C, Bartek J, Kallioniemi OP and Nevanlinna H . (2002). Am. J. Hum. Genet., 71, 432–438.
Wu XL, Webster SR and Chen JJ . (2001). J. Biol. Chem., 276, 2971–2974.
Xu XZ, Tsvetkov LA and Stern DF . (2002). Mol. Cell. Biol., 22, 4419–4432.
Zheng YX, Li L, Shen HB, Sturgis EM, Eicher SA, Strom SS, Spitz MR and Wei QY . (2001). Carcinogenesis, 22, 2005–2008.
Zhou BB and Bartek J . (2004). Nat. Rev. Cancer, 4, 216–225.
Acknowledgements
This work was supported by grants from the Norwegian Cancer Society, Nordic Cancer Union, Danish Cancer Society, and the European Union.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Staalesen, V., Falck, J., Geisler, S. et al. Alternative splicing and mutation status of CHEK2 in stage III breast cancer. Oncogene 23, 8535–8544 (2004). https://doi.org/10.1038/sj.onc.1207928
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1207928
Keywords
This article is cited by
-
Acetylation dependent translocation of EWSR1 regulates CHK2 alternative splicing in response to DNA damage
Oncogene (2022)
-
High PTEN gene expression is a negative prognostic marker in human primary breast cancers with preserved p53 function
Breast Cancer Research and Treatment (2017)
-
Prevalence of the CHEK2 R95* germline mutation
Hereditary Cancer in Clinical Practice (2016)
-
Genomic profiling of CHEK2*1100delC-mutated breast carcinomas
BMC Cancer (2015)
-
MicroRNA-191 promotes osteosarcoma cells proliferation by targeting checkpoint kinase 2
Tumor Biology (2015)