Identification and characterization of novel SNPs in CHEK2 in Ashkenazi Jewish men with prostate cancer

Abstract

Checkpoint kinase 2 (CHEK2) is a protein involved in arresting cell cycle in response to DNA damage. To investigate whether it plays an important role in the development of prostate cancer (PRCA) in the Ashkenazi Jewish (AJ) population, we sequenced CHEK2 in 75 AJ individuals with prostate, breast, or no cancer (n=25 each). We identified seven coding SNPs (five are novel) that changed the amino-acid sequence, resulting in R3W, E394F, Y424H, S428F, D438Y, P509S, and P509L. We determined the frequency of each variant in 76 AJ families collected by members of the International Consortium for Prostate Cancer Genetics (ICPCG) where >or=2 men were affected by PRCA. Only one variant, Y424H in exon 11, was identified in more than two families. Exon 11 was then screened in nine additional AJ ICPCG families (a total of 85 families). The Y424H variant occurred in nine affected cases from four different families; however, it did not completely segregate with the disease. We performed bioinformatics analysis, which showed that Y424H is a non-conservative missense substitution that falls at a position that is invariant in vertebrate CHEK2 orthologs. Both SIFT and Align-GVGD predict that Y424H is a loss of function mutation. However, the frequency of Y424H was not significantly different between unselected AJ cases from Montreal/Memorial Sloan Kettering Cancer Centre (MSKCC) and AJ controls from Israel/MSKCC (OR 1.18, 95%CI: 0.34-4.61, p=.99). Moreover, functional assays using Saccharomyces cerevisiae revealed that the Y424H substitution did not alter function of CHEK2 protein. Although we cannot rule out a subtle influence of the CHEK2 variants on PRCA risk, these results suggest that germline CHEK2 mutations have a minor role in, if any, PRCA susceptibility in AJ men.

FIGURE 1

Transformation with plasmids carrying wild type (wt) human CHEK2, as well as the Y424H variant, represses the normally reduced viability due to Rad53 deletion in the Rad53-null sml1-null yeast S. Cerevisae. Yeast carrying the CHEK2*1100delC variant grows less than that carrying the wild type, while yeast cells carrying the CHEK2*S428F variant grow more than cells carrying 1100delC but less than those carrying the wild type. Yeast carrying F428S, a mutant created from the S428F plasmid to re-create a wild type CHEK2 plasmid, grows as well as yeast carrying genuine wild type CHEK2. Time = time of measurement after inoculation of known number of yeast cells into fresh medium.