A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor
<i>Saccharomyces cerevisiae</i> does not contain a p53 homolog. Utilizing this yeast as an in vivo test tube model, our aim was to investigate if a yeast protein would show p53 DNA binding homology. Electrophoretic mobility shift analyses revealed the formation of specific DNA-protein co...
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| Format: | Article |
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MDPI AG
2020-03-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/3/417 |
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| author | Kanwal Farooqi Marjan Ghazvini Leah D. Pride Louis Mazzella David White Ajay Pramanik Jill Bargonetti Carol Wood Moore |
| author_facet | Kanwal Farooqi Marjan Ghazvini Leah D. Pride Louis Mazzella David White Ajay Pramanik Jill Bargonetti Carol Wood Moore |
| author_sort | Kanwal Farooqi |
| collection | DOAJ |
| description | <i>Saccharomyces cerevisiae</i> does not contain a p53 homolog. Utilizing this yeast as an in vivo test tube model, our aim was to investigate if a yeast protein would show p53 DNA binding homology. Electrophoretic mobility shift analyses revealed the formation of specific DNA-protein complexes consisting of <i>S. cerevisiae</i> nuclear protein(s) and oligonucleotides containing p53 DNA binding sites. A <i>S. cerevisiae</i> p53 binding site factor (Scp53BSF) bound to a p53 synthetic DNA-consensus sequence (SCS) and a p53 binding-site sequence from the <i>MDM2</i> oncogene. The complexes were of comparable size. Like mammalian p53, the affinity of Scp53BSF for the SCS oligonucleotide was higher than for the MDM2 oligonucleotide. Binding of Scp53BSF to the SCS and MDM2 oligonucleotides was strongly competed by unlabeled oligonucleotides containing mammalian p53 sites, but very little by a mutated site oligonucleotide. Importantly, Scp53BSF-DNA binding activity was significantly induced in extracts from cells with DNA damage. This resulted in dose-dependent coordinated activation of transcription when using p53-binding site reporter constructs. An ancient p53-like DNA binding protein may have been found, and activation of DNA-associated factors to p53 response elements may have functions not yet determined. |
| first_indexed | 2024-12-13T03:05:18Z |
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| id | doaj.art-51e65df38dae4e4992c29fef7969a748 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-12-13T03:05:18Z |
| publishDate | 2020-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-51e65df38dae4e4992c29fef7969a7482022-12-22T00:01:44ZengMDPI AGBiomolecules2218-273X2020-03-0110341710.3390/biom10030417biom10030417A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor SuppressorKanwal Farooqi0Marjan Ghazvini1Leah D. Pride2Louis Mazzella3David White4Ajay Pramanik5Jill Bargonetti6Carol Wood Moore7Department of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USADepartment of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USADepartment of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USADepartment of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USACity University of New York Graduate Center, Programs in Biochemistry and Biology, 365 Fifth Ave, New York, NY 10016, USADepartment of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USACity University of New York Graduate Center, Programs in Biochemistry and Biology, 365 Fifth Ave, New York, NY 10016, USADepartment of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USA<i>Saccharomyces cerevisiae</i> does not contain a p53 homolog. Utilizing this yeast as an in vivo test tube model, our aim was to investigate if a yeast protein would show p53 DNA binding homology. Electrophoretic mobility shift analyses revealed the formation of specific DNA-protein complexes consisting of <i>S. cerevisiae</i> nuclear protein(s) and oligonucleotides containing p53 DNA binding sites. A <i>S. cerevisiae</i> p53 binding site factor (Scp53BSF) bound to a p53 synthetic DNA-consensus sequence (SCS) and a p53 binding-site sequence from the <i>MDM2</i> oncogene. The complexes were of comparable size. Like mammalian p53, the affinity of Scp53BSF for the SCS oligonucleotide was higher than for the MDM2 oligonucleotide. Binding of Scp53BSF to the SCS and MDM2 oligonucleotides was strongly competed by unlabeled oligonucleotides containing mammalian p53 sites, but very little by a mutated site oligonucleotide. Importantly, Scp53BSF-DNA binding activity was significantly induced in extracts from cells with DNA damage. This resulted in dose-dependent coordinated activation of transcription when using p53-binding site reporter constructs. An ancient p53-like DNA binding protein may have been found, and activation of DNA-associated factors to p53 response elements may have functions not yet determined.https://www.mdpi.com/2218-273X/10/3/417p53 response elementsyeastscsmdm2transactivationp53 dna binding sitesp53 binding site factor<i>saccharomyes cerevisiae</i> |
| spellingShingle | Kanwal Farooqi Marjan Ghazvini Leah D. Pride Louis Mazzella David White Ajay Pramanik Jill Bargonetti Carol Wood Moore A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor Biomolecules p53 response elements yeast scs mdm2 transactivation p53 dna binding sites p53 binding site factor <i>saccharomyes cerevisiae</i> |
| title | A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor |
| title_full | A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor |
| title_fullStr | A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor |
| title_full_unstemmed | A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor |
| title_short | A Protein in the Yeast <i>Saccharomyces cerevisiae</i> Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor |
| title_sort | protein in the yeast i saccharomyces cerevisiae i presents dna binding homology to the p53 checkpoint protein and tumor suppressor |
| topic | p53 response elements yeast scs mdm2 transactivation p53 dna binding sites p53 binding site factor <i>saccharomyes cerevisiae</i> |
| url | https://www.mdpi.com/2218-273X/10/3/417 |
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