The p53 protein is a key tumor suppressor and the most commonly mutated and down-regulated protein in human tumors. It functions mainly through interaction with DNA, and p53 acts as a transcription factor that recognizes the so-called p53 target sites on the promoters of various genes. P53 has been shown to exist as many isoforms, including three C-terminal isoforms that are produced by alternative splicing. Because the C-terminal domain is responsible for sequence-nonspecific binding and regulation of p53 binding, we have analyzed DNA recognition by these C-terminal isoforms. Using atomic force microscopy, we show for the first time that all C-terminal isoforms recognize superhelical DNA. It is particularly noteworthy that a sequence-specific p53 consensus binding site is bound by p53α and β isoforms with similar affinities, whilst p53α shows higher binding to a quadruplex sequence than both p53β and p53γ, and p53γ loses preferential binding to both the consensus binding sequence and the quadruplex-forming sequence. These results show the important role of the variable p53 C-terminal amino acid sequences for DNA recognition.

Department of Informatics Mendel University in Brno Zemědělská 1 613 00 Brno Czech Republic

Faculty of Chemistry Brno University of Technology Purkyňova 118 612 00 Brno Czech Republic

Faculty of Mechanical Engineering Brno University of Technology Technická 2 616 69 Brno Czech Republic

Institute of Biophysics of the Czech Academy of Sciences Královopolská 135 612 00 Brno Czech Republic

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$a Interaction of C-terminal p53 isoforms depends strongly upon DNA sequence and topology / $c P. Goswami, L. Šislerová, M. Dobrovolná, J. Havlík, J. Šťastný, V. Brázda

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$a The p53 protein is a key tumor suppressor and the most commonly mutated and down-regulated protein in human tumors. It functions mainly through interaction with DNA, and p53 acts as a transcription factor that recognizes the so-called p53 target sites on the promoters of various genes. P53 has been shown to exist as many isoforms, including three C-terminal isoforms that are produced by alternative splicing. Because the C-terminal domain is responsible for sequence-nonspecific binding and regulation of p53 binding, we have analyzed DNA recognition by these C-terminal isoforms. Using atomic force microscopy, we show for the first time that all C-terminal isoforms recognize superhelical DNA. It is particularly noteworthy that a sequence-specific p53 consensus binding site is bound by p53α and β isoforms with similar affinities, whilst p53α shows higher binding to a quadruplex sequence than both p53β and p53γ, and p53γ loses preferential binding to both the consensus binding sequence and the quadruplex-forming sequence. These results show the important role of the variable p53 C-terminal amino acid sequences for DNA recognition.

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$a Šislerová, Lucie $u Institute of Biophysics of the Czech Academy of Sciences, Královopolská 135, 612 00, Brno, Czech Republic; Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 612 00, Brno, Czech Republic

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$a Dobrovolná, Michaela $u Institute of Biophysics of the Czech Academy of Sciences, Královopolská 135, 612 00, Brno, Czech Republic; Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 612 00, Brno, Czech Republic

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$a Havlík, Jan $u Department of Informatics, Mendel University in Brno, Zemědělská 1, 613 00, Brno, Czech Republic

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$a Šťastný, Jiří $u Department of Informatics, Mendel University in Brno, Zemědělská 1, 613 00, Brno, Czech Republic; Faculty of Mechanical Engineering, Brno University of Technology, Technická 2, 616 69, Brno, Czech Republic

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$a Brázda, Václav $u Institute of Biophysics of the Czech Academy of Sciences, Královopolská 135, 612 00, Brno, Czech Republic; Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 612 00, Brno, Czech Republic. Electronic address: vaclav@ibp.cz

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