G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter
The regulation of conformational arrangements of gene promoters is a physiological mechanism that has been associated with the fine control of gene expression. Indeed, it can drive the time and the location for the selective recruitment of proteins of the transcriptional machinery. Here, we address...
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| Format: | Article |
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MDPI AG
2020-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/1/329 |
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| author | Silvia Da Ros Giulia Nicoletto Riccardo Rigo Silvia Ceschi Eleonora Zorzan Mauro Dacasto Mery Giantin Claudia Sissi |
| author_facet | Silvia Da Ros Giulia Nicoletto Riccardo Rigo Silvia Ceschi Eleonora Zorzan Mauro Dacasto Mery Giantin Claudia Sissi |
| author_sort | Silvia Da Ros |
| collection | DOAJ |
| description | The regulation of conformational arrangements of gene promoters is a physiological mechanism that has been associated with the fine control of gene expression. Indeed, it can drive the time and the location for the selective recruitment of proteins of the transcriptional machinery. Here, we address this issue at the <i>KIT</i> proximal promoter where three G-quadruplex forming sites are present (kit1, kit2 and kit*). On this model, we focused on the interplay between G-quadruplex (G4) formation and SP1 recruitment. By site directed mutagenesis, we prepared a library of plasmids containing mutated sequences of the WT <i>KIT</i> promoter that systematically exploited different G4 formation attitudes and SP1 binding properties. Our transfection data showed that the three different G4 sites of the <i>KIT</i> promoter impact on SP1 binding and protein expression at different levels. Notably, kit2 and kit* structural features represent an on-off system for <i>KIT</i> expression through the recruitment of transcription factors. The use of two G4 binders further helps to address kit2-kit* as a reliable target for pharmacological intervention. |
| first_indexed | 2024-03-10T13:38:41Z |
| format | Article |
| id | doaj.art-635fe677dcd64618aa409350e23683f0 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T13:38:41Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-635fe677dcd64618aa409350e23683f02023-11-21T03:12:29ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-0122132910.3390/ijms22010329G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal PromoterSilvia Da Ros0Giulia Nicoletto1Riccardo Rigo2Silvia Ceschi3Eleonora Zorzan4Mauro Dacasto5Mery Giantin6Claudia Sissi7Department of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, ItalyDepartment of Comparative Biomedicine and Food Science, University of Padua, 35020 Legnaro, ItalyDepartment of Comparative Biomedicine and Food Science, University of Padua, 35020 Legnaro, ItalyDepartment of Comparative Biomedicine and Food Science, University of Padua, 35020 Legnaro, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, 35131 Padua, ItalyThe regulation of conformational arrangements of gene promoters is a physiological mechanism that has been associated with the fine control of gene expression. Indeed, it can drive the time and the location for the selective recruitment of proteins of the transcriptional machinery. Here, we address this issue at the <i>KIT</i> proximal promoter where three G-quadruplex forming sites are present (kit1, kit2 and kit*). On this model, we focused on the interplay between G-quadruplex (G4) formation and SP1 recruitment. By site directed mutagenesis, we prepared a library of plasmids containing mutated sequences of the WT <i>KIT</i> promoter that systematically exploited different G4 formation attitudes and SP1 binding properties. Our transfection data showed that the three different G4 sites of the <i>KIT</i> promoter impact on SP1 binding and protein expression at different levels. Notably, kit2 and kit* structural features represent an on-off system for <i>KIT</i> expression through the recruitment of transcription factors. The use of two G4 binders further helps to address kit2-kit* as a reliable target for pharmacological intervention.https://www.mdpi.com/1422-0067/22/1/329G-quadruplex<i>KIT</i> promoterSP1gene expression |
| spellingShingle | Silvia Da Ros Giulia Nicoletto Riccardo Rigo Silvia Ceschi Eleonora Zorzan Mauro Dacasto Mery Giantin Claudia Sissi G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter International Journal of Molecular Sciences G-quadruplex <i>KIT</i> promoter SP1 gene expression |
| title | G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter |
| title_full | G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter |
| title_fullStr | G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter |
| title_full_unstemmed | G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter |
| title_short | G-Quadruplex Modulation of SP1 Functional Binding Sites at the <i>KIT</i> Proximal Promoter |
| title_sort | g quadruplex modulation of sp1 functional binding sites at the i kit i proximal promoter |
| topic | G-quadruplex <i>KIT</i> promoter SP1 gene expression |
| url | https://www.mdpi.com/1422-0067/22/1/329 |
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