Analysis of the DNA-binding activity of p53 mutants using functional protein microarrays and its relationship to transcriptional activation
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Jitka Malcikova
Abstract
Sequence-specific DNA binding is the key function through which tumor suppressor p53 exerts transactivation of the downstream target genes, often being impaired in cancer cells by mutations in the TP53 gene. Functional protein microarray technology enables a high-throughput parallel analysis of protein properties within one experiment under the same conditions. Using an array approach, we analyzed the DNA binding activity of wild type p53 protein and of 49 variants. Our results show significant differences in the binding properties between the p53 mutants. The C-terminal mutant R337C displayed the highest DNA binding activity on the array. However, the same mutant showed only a partial activation in the reporter gene assay and almost no activation of downstream target genes after transfection of expression vector into cells lacking endogenous p53. These observations demonstrate that DNA binding itself is not sufficient for activating the p53 target genes in at least some of the p53 mutants and, therefore, in vitro studies might not always reflect in vivo conditions.
©2010 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- EDITORIAL
- Highlight: Signal Transduction and Disease
- HIGHLIGHT: SIGNAL TRANSDUCTION AND DISEASE
- Phosphorylase and the origin of reversible protein phosphorylation
- β-Catenin as a multilayer modulator of zonal cytochrome P450 expression in mouse liver
- Glycome profiling using modern glycomics technology: technical aspects and applications
- Ubiquitin ligase complexes: from substrate selectivity to conjugational specificity
- MAP3K1 functionally interacts with Axin1 in the canonical Wnt signalling pathway
- Signal transduction by the atopy-associated human thymic stromal lymphopoietin (TSLP) receptor depends on Janus kinase function
- GENES AND NUCLEIC ACIDS
- 6S RNA-dependent inhibition of RNA polymerase is released by RNA-dependent synthesis of small de novo products
- PROTEIN STRUCTURE AND FUNCTION
- Analysis of the DNA-binding activity of p53 mutants using functional protein microarrays and its relationship to transcriptional activation
- Peptides from the Mycobacterium tuberculosis Rv1980c protein involved in human cell infection: insights into new synthetic subunit vaccine candidates
- The epimerase activity of anthocyanidin reductase from Vitis vinifera and its regiospecific hydride transfers
- Antibacterial activity of radical scavengers against class Ib ribonucleotide reductase from Bacillus anthracis
- CELL BIOLOGY AND SIGNALING
- Signal transduction in CHO cells stably transfected with domain-selective forms of murine ACE
- The impact of methylmercury on 1,25-dihydroxyvitamin D3-induced transcriptomic responses in dolphin skin cells
- PROTEOLYSIS
- An examination of the proteolytic activity for bovine pregnancy-associated glycoproteins 2 and 12
- Analysis of an autoproteolytic activity of rice yellow mottle virus silencing suppressor P1
- Identification of novel peptide inhibitors for human trypsins
