Figure 12
Nucleotide substitutions of P2RY5 in sporadic and hereditary cancers. (a) Summary of sequence analysis of P2RY5. The positions of nucleotide substitutions are shown on the full-length mRNA. Details of sequencing are provided in Supplementary Table 2. (b) A G-T polymorphism at codon 307 in the PBDNA resulting in substitution of cysteine for tryptophan was identified by pyrosequencing (same case as shown in (d)). (c) A model of inactive P2RY5 containing seven transmembrane (H1–H7) and one cytoplasmic (H8) helix structures showing the position of polymorphism in codon 307 located within the cytoplasmic domain of the protein (left diagram) that may affect its interaction with the Gαβγ trimeric protein complex (right diagram). (d) Pedigree of a family affected by several common human malignancies that include cancers of the breast, lung, colon, prostate, and uterus as well as acute leukemia. Sequencing of the peripheral blood DNA in individual IV1 identified a mutation of P2RY5. (e) Wild-type sequence of P2RY5 (upper panel). A missense G-C mutation involving codon 111 and resulting in substitution of threonine for serine (S → T) documented by sequencing with subcloning of peripheral blood DNA from individual IV1 shown in (d) (lower panel). (f) Confirmation of a missense G-C mutation involving codon 111 of P2RY5 in individuals IV1, IV6, and IV20 by pyrosequencing (same family as shown in (d)). Note a loss of wild-type P2RY5 allele and retention of a mutant P2RY5 in breast cancer cells from individual IV1. PBDNA, peripheral blood DNA; Br Ca DNA, DNA extracted from breast cancer cells microdissected with laser from paraffin-embedded tissue.
