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 allelespecific 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 atrisk patients.
| Original language | English |
|---|---|
| Pages (from-to) | 6387-6394 |
| Number of pages | 8 |
| Journal | Oncogene |
| Volume | 22 |
| Issue number | 41 |
| DOIs | |
| State | Published - Sep 25 2003 |
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All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cancer Research
- Genetics
Cite this
Germline splicing mutations of CDKN2A predispose to melanoma. / Loo, Joanne C Y; Liu, Ling; Hao, AiHua; Gao, LuZhuang; Agatep, Ron; Shennan, Michael; Summers, Anne; Goldstein, Alisa M.; Tucker, Margaret A.; Deters, Carolyn; Fusaro, Ramon; Blazer, Kathleen; Weitzel, Jeffrey; Lassam, Norman; Lynch, Henry T.; Hogg, David.
In: Oncogene, Vol. 22, No. 41, 25.09.2003, p. 6387-6394.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Germline splicing mutations of CDKN2A predispose to melanoma
AU - Loo, Joanne C Y
AU - Liu, Ling
AU - Hao, AiHua
AU - Gao, LuZhuang
AU - Agatep, Ron
AU - Shennan, Michael
AU - Summers, Anne
AU - Goldstein, Alisa M.
AU - Tucker, Margaret A.
AU - Deters, Carolyn
AU - Fusaro, Ramon
AU - Blazer, Kathleen
AU - Weitzel, Jeffrey
AU - Lassam, Norman
AU - Lynch, Henry T.
AU - Hogg, David
PY - 2003/9/25
Y1 - 2003/9/25
N2 - 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 allelespecific 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 atrisk patients.
AB - 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 allelespecific 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 atrisk patients.
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UR - http://www.scopus.com/inward/citedby.url?scp=0142072167&partnerID=8YFLogxK
U2 - 10.1038/sj.onc.1206736
DO - 10.1038/sj.onc.1206736
M3 - Article
C2 - 14508519
AN - SCOPUS:0142072167
VL - 22
SP - 6387
EP - 6394
JO - Oncogene
JF - Oncogene
SN - 0950-9232
IS - 41
ER -