<i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation
Occurrence of non-canonical G-quadruplex (G4) DNA structures in the genome have been recognized as key factors in gene regulation and several other cellular processes. The <i>mosR</i> and <i>ndhA genes</i> involved in pathways of oxidation sensing regulation and ATP generatio...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/14/5/978 |
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| author | Arpita Dey Kushi Anand Amit Singh Ramasare Prasad Ritu Barthwal |
| author_facet | Arpita Dey Kushi Anand Amit Singh Ramasare Prasad Ritu Barthwal |
| author_sort | Arpita Dey |
| collection | DOAJ |
| description | Occurrence of non-canonical G-quadruplex (G4) DNA structures in the genome have been recognized as key factors in gene regulation and several other cellular processes. The <i>mosR</i> and <i>ndhA genes</i> involved in pathways of oxidation sensing regulation and ATP generation, respectively, make <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) bacteria responsible for oxidative stress inside host macrophage cells. Circular Dichroism spectra demonstrate stable hybrid G4 DNA conformations of <i>mosR</i>/<i>ndhA</i> DNA sequences. Real-time binding of mitoxantrone to G4 DNA with an affinity constant ~10<sup>5</sup>–10<sup>7</sup> M<sup>−1</sup>, leads to hypochromism with a red shift of ~18 nm, followed by hyperchromism in the absorption spectra. The corresponding fluorescence is quenched with a red shift ~15 nm followed by an increase in intensity. A change in conformation of the G4 DNA accompanies the formation of multiple stoichiometric complexes with a dual binding mode. The external binding of mitoxantrone with a partial stacking with G-quartets and/or groove binding induces significant thermal stabilization, ~20–29 °C in <i>ndhA/mosR</i> G4 DNA. The interaction leads to a two/four-fold downregulation of transcriptomes of <i>mosR</i>/<i>ndhA</i> genes apart from the suppression of DNA replication by <i>Taq</i> polymerase enzyme, establishing the role of mitoxantrone in targeting G4 DNA, as an alternate strategy for effective anti-tuberculosis action in view of deadly multi-drug resistant tuberculosis disease causing bacterial strains t that arise from existing therapeutic treatments. |
| first_indexed | 2024-03-11T03:42:21Z |
| format | Article |
| id | doaj.art-2569170e7b594145918e390d1e48f87b |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-11T03:42:21Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-2569170e7b594145918e390d1e48f87b2023-11-18T01:28:48ZengMDPI AGGenes2073-44252023-04-0114597810.3390/genes14050978<i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene RegulationArpita Dey0Kushi Anand1Amit Singh2Ramasare Prasad3Ritu Barthwal4Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee 247667, Uttarakhand, IndiaCentre for Infectious Disease Research, Indian Institute of Science, Bengaluru 560012, Karnataka, IndiaCentre for Infectious Disease Research, Indian Institute of Science, Bengaluru 560012, Karnataka, IndiaDepartment of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee 247667, Uttarakhand, IndiaDepartment of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee 247667, Uttarakhand, IndiaOccurrence of non-canonical G-quadruplex (G4) DNA structures in the genome have been recognized as key factors in gene regulation and several other cellular processes. The <i>mosR</i> and <i>ndhA genes</i> involved in pathways of oxidation sensing regulation and ATP generation, respectively, make <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) bacteria responsible for oxidative stress inside host macrophage cells. Circular Dichroism spectra demonstrate stable hybrid G4 DNA conformations of <i>mosR</i>/<i>ndhA</i> DNA sequences. Real-time binding of mitoxantrone to G4 DNA with an affinity constant ~10<sup>5</sup>–10<sup>7</sup> M<sup>−1</sup>, leads to hypochromism with a red shift of ~18 nm, followed by hyperchromism in the absorption spectra. The corresponding fluorescence is quenched with a red shift ~15 nm followed by an increase in intensity. A change in conformation of the G4 DNA accompanies the formation of multiple stoichiometric complexes with a dual binding mode. The external binding of mitoxantrone with a partial stacking with G-quartets and/or groove binding induces significant thermal stabilization, ~20–29 °C in <i>ndhA/mosR</i> G4 DNA. The interaction leads to a two/four-fold downregulation of transcriptomes of <i>mosR</i>/<i>ndhA</i> genes apart from the suppression of DNA replication by <i>Taq</i> polymerase enzyme, establishing the role of mitoxantrone in targeting G4 DNA, as an alternate strategy for effective anti-tuberculosis action in view of deadly multi-drug resistant tuberculosis disease causing bacterial strains t that arise from existing therapeutic treatments.https://www.mdpi.com/2073-4425/14/5/978G-quadruplex<i>mosR</i>/<i>ndhA</i> genesdownregulationDNA replicationDNA targeting by mitoxantronespectroscopy and calorimetry |
| spellingShingle | Arpita Dey Kushi Anand Amit Singh Ramasare Prasad Ritu Barthwal <i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation Genes G-quadruplex <i>mosR</i>/<i>ndhA</i> genes downregulation DNA replication DNA targeting by mitoxantrone spectroscopy and calorimetry |
| title | <i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation |
| title_full | <i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation |
| title_fullStr | <i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation |
| title_full_unstemmed | <i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation |
| title_short | <i>MOSR</i> and <i>NDH</i>A Genes Comprising G-Quadruplex as Promising Therapeutic Targets against <i>Mycobacterium tuberculosis</i>: Molecular Recognition by Mitoxantrone Suppresses Replication and Gene Regulation |
| title_sort | i mosr i and i ndh i a genes comprising g quadruplex as promising therapeutic targets against i mycobacterium tuberculosis i molecular recognition by mitoxantrone suppresses replication and gene regulation |
| topic | G-quadruplex <i>mosR</i>/<i>ndhA</i> genes downregulation DNA replication DNA targeting by mitoxantrone spectroscopy and calorimetry |
| url | https://www.mdpi.com/2073-4425/14/5/978 |
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