Frequent constitutional C to T mutations in CGA-arginine codons in the RB1 gene produce premature stop codons in patients with bilateral (hereditary) retinoblastoma.

We have shown previously that the most common point mutation in the RB1 gene in retinoblastoma tumours is a C-->T transition and that the majority of these occur in CGAarg codons. As a result of this mutation, a TGAstop codon is generated directly. We have analysed the 14 CGAarg codons in the RB1 gene for mutations in 113 patients with bilateral retinoblastoma. At 6 of these sites, C-->T mutations in CGA codons alter a restriction enzyme site which makes their identification relatively straightforward. It was necessary, however, to analyse the other 8 CGA codons using single-strand conformation polymorphism (SSCP) analysis. A total of 18 C-->T mutations were found, which represents 16% of all patients. Of these 13 (73%) were at two particular CGA codons in exon 8 (codon 251) and exon 17 (codon 552). During the course of the SSCP analysis, mutations were identified in 7 other individuals. Thus, 20-25% of all mutations can be identified by a relatively quick survey of the CGA codons in the RB1 gene, which has important implications for genetic screening programmes. All of the mutations in the RB1 gene in these bilaterally affected patients would be predicted to result in the absence of a functional protein.