Abstract
Approximately 50% of all melanoma families worldwide show linkage to 9p21‐22, but only about half of these have been shown to contain germ line CDKN2A mutations. It has been hypothesized that a proportion of these families carry mutations in the noncoding regions of CDKN2A. Several Canadian families have been reported to carry a mutation in the 5′ UTR, at position −34 relative to the start site, which gives rise to a novel AUG translation initiation codon that markedly decreases translation from the wild‐type AUG (Liu et al., 1999). Haplotype sharing in these Canadian families suggested that this mutation is of British origin. We sequenced 1,327 base pairs (bp) of CDKN2A, making up 1,116 bp of the 5′ UTR and promoter, all of exon 1, and 61 bp of intron 1, in at least one melanoma case from 110 Australian families with three or more affected members known not to carry mutations within the p16 coding region. In addition, 431 bp upstream of the start codon was sequenced in an additional 253 affected probands from two‐case melanoma families for which the CDKN2A mutation status was unknown. Several known polymorphisms at positions −33, −191, −493, and −735 were detected, in addition to four novel variants at positions 120, −252, −347, and −981 relative to the start codon. One of the probands from a two‐case family was found to have the previously reported Q50R mutation. No family member was found to carry the mutation at position −34 or any other disease‐associated mutation. For further investigation of noncoding CDKN2A mutations that may affect transcription, allele‐specific expression analysis was carried out in 31 of the families with at least three affected members who showed either complete or “indeterminate” 9p haplotype sharing without CDKN2A exonic mutations. Reverse transcription polymerase chain reaction and automated sequencing showed expression of both CDKN2A alleles in all family members tested. The lack of CDKN2A promoter mutations and the absence of transcriptional silencing in the germ line of this cohort of families suggest that mutations in the promoter and 5′ UTR play a very limited role in melanoma predisposition. © 2001 Wiley‐Liss, Inc.