MALČÍKOVÁ, Jitka, Boris TICHÝ, Jiří DAMBORSKÝ, Jitka KABÁTHOVÁ, Martin TRBUŠEK, Jiří MAYER a Šárka POSPÍŠILOVÁ. Analysis of the DNA-binding activity of p53 mutants using functional protein microarrays and its relationship to transcriptional activation. Biological Chemistry. Berlin, DE: Walter de Gruyter&CO, 2010, roč. 391, 2/3, s. 197-205. ISSN 1431-6730. |
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@article{879295, author = {Malčíková, Jitka and Tichý, Boris and Damborský, Jiří and Kabáthová, Jitka and Trbušek, Martin and Mayer, Jiří and Pospíšilová, Šárka}, article_location = {Berlin, DE}, article_number = {2/3}, keywords = {R337C; real-time PCR; reporter gene assay; transactivation}, language = {eng}, issn = {1431-6730}, journal = {Biological Chemistry}, title = {Analysis of the DNA-binding activity of p53 mutants using functional protein microarrays and its relationship to transcriptional activation}, volume = {391}, year = {2010} }
TY - JOUR ID - 879295 AU - Malčíková, Jitka - Tichý, Boris - Damborský, Jiří - Kabáthová, Jitka - Trbušek, Martin - Mayer, Jiří - Pospíšilová, Šárka PY - 2010 TI - Analysis of the DNA-binding activity of p53 mutants using functional protein microarrays and its relationship to transcriptional activation JF - Biological Chemistry VL - 391 IS - 2/3 SP - 197-205 EP - 197-205 PB - Walter de Gruyter&CO SN - 14316730 KW - R337C KW - real-time PCR KW - reporter gene assay KW - transactivation N2 - 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. ER -
MALČÍKOVÁ, Jitka, Boris TICHÝ, Jiří DAMBORSKÝ, Jitka KABÁTHOVÁ, Martin TRBUŠEK, Jiří MAYER a Šárka POSPÍŠILOVÁ. Analysis of the DNA-binding activity of p53 mutants using functional protein microarrays and its relationship to transcriptional activation. \textit{Biological Chemistry}. Berlin, DE: Walter de Gruyter\&{}CO, 2010, roč.~391, 2/3, s.~197-205. ISSN~1431-6730.
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