Abstract
Coding mutations of the CDKN2A gene on chromosome 9p21 cosegregate with 25–60% of familial melanoma cases, but there remains a number of 9p21-linked kindreds that lack germline coding mutations of CDKN2A. We sequenced CDKN2A exons 1α, 2, 3, and the adjacent intronic regions in 167 melanoma-prone families (at least two affected first-degree relatives), and detected four splice site variations, three of which cosegregate with the disease. RT–PCR experiments verified that these three variants, including an AGgt to ATgt mutation that demonstrates a founder effect, do affect splicing. While an exon 1α splice donor site mutation incompletely abolishes splicing, the correctly spliced mRNA yields a protein (Q50P) that cannot effectively interact with CDK4. We also performed RT–PCR on mRNA from 16 melanoma-prone kindreds to search for cryptic splice sites deep within introns, but identified no splice variants. Meanwhile, we screened 139 affected families using allele-specific PCR for the recently discovered IVS2−105A>G mutation, but found only one family that possesses this alteration. We conclude that splice site mutations do predispose to disease in a subset of melanoma-prone kindreds. Characterization of additional splice site variants and other noncoding alterations of CDKN2A should allow us to detect a wider range of mutations in at-risk patients.
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References
Byeon IJ, Li J, Ericson K, Selby TL, Tevelev A, Kim HJ, O'Maille P and Tsai MD . (1998). Mol. Cell, 1, 421–431.
Ciotti P, Struewing JP, Mantelli M, Chompret A, Avril MF, Santi PL, Tucker MA, Bianchi-Scarra G, Bressac-de Paillerets B and Goldstein AM . (2000). Am. J. Hum. Genet., 67, 311–319.
Coligan J, Kruisbeek A, Margulies D, Shevach E and Strober W . (1999). Current Protocols in Immunology, Vol. Unit 7.22.
Goldstein AM, Liu L, Shennan MG, Hogg D, Tucker MA and Struewing JP . (2001). Br. J. Cancer, 85, 527–530.
Gruis NA, Sandkuijl LA, van der Velden PA, Bergman W and Frants RR . (1995). Melanoma Res., 5, 169–177.
Harland M, Mistry S, Bishop DT and Bishop JA . (2001). Hum. Mol. Genet., 10, 2679–2686.
Hashemi JB, Bendahl PO, Sandberg T, Platz A, Linder S, Stierner U, Olsson H, Ingvar C, Hansson J and Borg A . (2001). Genes Chromosomes Cancer, 31, 107–116.
Hewitt C, Wu CL, Evans G, Howell A, Elles RG, Jordan R, Sloan P, Read AP and Thakker N . (2002). Hum. Mol. Genet., 11, 1273–1279.
Hogg D, Brill H, Liu L, Monzon J, Summers A, From L and Lassam NJ . (1998). J. Cutan. Med. Surg., 2, 172–179.
Hussussian CJ, Struewing JP, Goldstein AM, Higgins PA, Ally DS, Sheahan MD, Clark Jr WH, Tucker MA and Dracopoli NC . (1994). Nat. Genet., 8, 15–21.
Li J, Byeon IJ, Ericson K, Poi MJ, O'Maille P, Selby T and Tsai MD . (1999). Biochemistry, 38, 2930–2940.
Liu L, Dilworth D, Gao L, Monzon J, Summers A, Lassam N and Hogg D . (1999). Nat. Genet., 21, 128–132.
Liu L, Goldstein AM, Tucker MA, Brill H, Gruis NA, Hogg D and Lassam NJ . (1997). Genes Chromosomes Cancer, 19, 52–54.
Liu L, Lassam NJ, Slingerland JM, Bailey D, Cole D, Jenkins R and Hogg D . (1995). Oncogene, 11, 405–412.
Luh FY, Archer SJ, Domaille PJ, Smith BO, Owen D, Brotherton DH, Raine AR, Xu X, Brizuela L, Brenner SL and Laue ED . (1997). Nature, 389, 999–1003.
Lynch HT, Brand RE, Hogg D, Deters CA, Fusaro RM, Lynch JF, Liu L, Knezetic J, Lassam NJ, Goggins M and Kern S . (2002). Cancer, 94, 84–96.
MacKie RM, Andrew N, Lanyon WG and Connor JM . (1998). J. Invest. Dermatol., 111, 269–272.
Monzon J, Liu L, Brill H, Goldstein AM, Tucker MA, From L, McLaughlin J, Hogg D and Lassam NJ . (1998). N. Engl. J. Med., 338, 879–887.
Moskaluk CA, Hruban H, Lietman A, Smyrk T, Fusaro L, Fusaro R, Lynch J, Yeo CJ, Jackson CE, Lynch HT and Kern SE . (1998). Hum. Mutat., 12, 70.
Petronzelli F, Sollima D, Coppola G, Martini-Neri ME, Neri G and Genuardi M . (2001). Genes Chromosomes Cancer, 31, 398–401.
Pollock PM, Spurr N, Bishop T, Newton-Bishop J, Gruis N, van der Velden PA, Goldstein AM, Tucker MA, Foulkes WD, Barnhill R, Haber D, Fountain J and Hayward NK . (1998). Hum. Mutat., 11, 424–431.
Quelle DE, Cheng M, Ashmun RA and Sherr CJ . (1997). Proc. Natl. Acad. Sci. USA, 94, 669–673.
Rizos H, Puig S, Badenas C, Malvehy J, Darmanian AP, Jiménez L, Milà M and Kefford RF . (2001). Oncogene, 20, 5543–5547.
Ruas M and Peters G . (1998). Biochim. Biophys. Acta, 1378, F115–F177.
Rutter JL, Goldstein AM, Davila MR, Tucker MA, Struewing JP . (in press). Oncogene.
Shapiro MB and Senapathy P . (1987). Nucleic Acids Res., 15, 7155–7174.
Shennan MG, Badin AC, Walsh S, Summers A, From L, McKenzie M, Goldstein AM, Tucker MA, Hogg D and Lassam N . (2000). Oncogene, 19, 1849–1852.
Acknowledgements
We thank the individuals and families who agreed to take part in these studies; S Hollenberg (Fred Hutchinson Cancer Research Center, Seattle) for yeast two-hybrid study reagents; and C Arrowsmith (Ontario Cancer Institute, Toronto) and other members of the Hogg and Lassam Laboratories for helpful discussions. JCYL and RA were supported by an Ontario Graduate Scholarship and the Canadian Institutes of Health Research KM Hunter Charitable Foundation Doctoral Research Award, respectively. JW and KB (City of Hope Cancer Center) were supported in part by California Cancer Research Program of the University of California, Grant Number 99–86874. This work was supported by the National Cancer Institute of Canada (NCIC).
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Dr Lassam is deceased.
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Loo, J., Liu, L., Hao, A. et al. Germline splicing mutations of CDKN2A predispose to melanoma. Oncogene 22, 6387–6394 (2003). https://doi.org/10.1038/sj.onc.1206736
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DOI: https://doi.org/10.1038/sj.onc.1206736
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