title: Functional protein microarrays for parallel characterisation of p53 mutants.
abstract: Understanding the way in which single nucleotide polymorphisms and mutations in the human genome result in individual susceptibility to disease is a major goal in the postgenomic era. Such knowledge should accelerate the development of personalised medicine in which drug treatment can specifically match an individual's genotype. High-throughput DNA sequencing is generating the initial information required, but new technologies are required that can rapidly characterise the phenotypic effects of the identified polymorphisms. For example, many thousands of allelic variants of the p53 gene have been described and are responsible for more than 50% of cancers, however few of the protein products have been functionally characterised. Here we have quantified in parallel the effects of mutations and polymorphisms on the DNA-binding function of the p53 oncoprotein using a protein microarray, allowing their subclassification according to functional effect. Protein-protein interactions between p53 variants and (i) a regulatory oncoprotein, (ii) a regulatory kinase resulting in on-chip phosphorylation, are also described, suggesting the more general utility of this high-throughput assay format.
authors.affiliation: Procognia Ltd., The Switchback, Gardner Road, Maidenhead SL6 7RJ, UK.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Boutell
authors.forename: Darren J
authors.initials: DJ
authors.lastname: Hart
authors.forename: Benjamin L J
authors.initials: BL
authors.lastname: Godber
authors.forename: Roland Z
authors.initials: RZ
authors.lastname: Kozlowski
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Blackburn
mesh_terms: D000483:Alleles; D047390:Casein Kinase II / Q000737:chemistry; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D005456:Fluorescent Dyes / Q000494:pharmacology; D016158:Genes, p53; D014644:Genetic Variation; D005822:Genetic Vectors; D005838:Genotype; D006801:Humans; D007700:Kinetics; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D010766:Phosphorylation; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2; D017422:Sequence Analysis, DNA; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D016159:Tumor Suppressor Protein p53; D004247:DNA; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D047390:Casein Kinase II