Portal de Periódicos da CAPES

Predicting functional significance of cancer-associated p16INK4a mutations in CDKN2A

2010; Wiley; Volume: 31; Issue: 6 Linguagem: Inglês

10.1002/humu.21245

1098-1004

Heather McKenzie, Carina Fung, Therese M. Becker, Mal Irvine, Graham J. Mann, Richard Kefford, Helen Rizos,

Cancer Immunotherapy and Biomarkers

Inherited mutations affecting the INK4a/ARF locus (CDKN2A) are associated with melanoma susceptibility in 40% of multiple case melanoma families. Over 60 different germline INK4a/ARF mutations have been detected in more than 190 families worldwide. The majority of these alterations are missense mutations affecting p16INK4a, and only 25% of these have been functionally assessed. There is therefore a need for an accurate and rapid assay to determine the functional significance of p16INK4a mutations. We reviewed the performance of several in vivo functional assays that measure critical aspects of p16INK4a function, including subcellular location, CDK binding and cell cycle inhibition. In this report the function of 28 p16INK4a variants, many associated with melanoma susceptibility were compared. We show that assessment of CDK4 binding and subcellular localization can accurately and rapidly determine the functional significance of melanoma-associated p16INK4a mutations. p16INK4a-CDK6 binding affinity was unhelpful, as no disease-associated mutation showed reduced CDK6 affinity while maintaining the ability to bind CDK4. Likewise, in silico analyses did not contribute substantially, with only 12 of 25 melanoma-associated missense variants consistently predicted as deleterious. The ability to determine variant functional activity accurately would identify disease-associated mutations and facilitate effective genetic counselling of individuals at high risk of melanoma. Hum Mutat 31:1–10, 2010. © 2010 Wiley-Liss, Inc.