Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation
ABSTRACT The human malaria parasite undergoes a noncanonical cell division, namely, endoreduplication, where several rounds of nuclear, mitochondrial, and apicoplast replication occur without cytoplasmic division. Despite its importance in Plasmodium biology, the topoisomerases essential for decaten...
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| Format: | Article |
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American Society for Microbiology
2023-06-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.04980-22 |
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| author | Priyanka Singh Wahida Tabassum Nupur Fangaria Sandeep Dey Siladitya Padhi Mrinal K. Bhattacharyya Kota Arun Kumar Arijit Roy Sunanda Bhattacharyya |
| author_facet | Priyanka Singh Wahida Tabassum Nupur Fangaria Sandeep Dey Siladitya Padhi Mrinal K. Bhattacharyya Kota Arun Kumar Arijit Roy Sunanda Bhattacharyya |
| author_sort | Priyanka Singh |
| collection | DOAJ |
| description | ABSTRACT The human malaria parasite undergoes a noncanonical cell division, namely, endoreduplication, where several rounds of nuclear, mitochondrial, and apicoplast replication occur without cytoplasmic division. Despite its importance in Plasmodium biology, the topoisomerases essential for decatenation of replicated chromosome during endoreduplication remain elusive. We hypothesize that the topoisomerase VI complex, containing Plasmodium falciparum topiosomerase VIB (PfTopoVIB) and catalytic P. falciparum Spo11 (PfSpo11), might be involved in the segregation of the Plasmodium mitochondrial genome. Here, we demonstrate that the putative PfSpo11 is the functional ortholog of yeast Spo11 that can complement the sporulation defects of the yeast Δspo11 strain, and the catalytic mutant Pfspo11Y65F cannot complement such defects. PfTopoVIB and PfSpo11 display a distinct expression pattern compared to the other type II topoisomerases of Plasmodium and are induced specifically at the late schizont stage of the parasite, when the mitochondrial genome segregation occurs. Furthermore, PfTopoVIB and PfSpo11 are physically associated with each other at the late schizont stage, and both subunits are localized in the mitochondria. Using PfTopoVIB- and PfSpo11-specific antibodies, we immunoprecipitated the chromatin of tightly synchronous early, mid-, and late schizont stage-specific parasites and found that both the subunits are associated with the mitochondrial genome during the late schizont stage of the parasite. Furthermore, PfTopoVIB inhibitor radicicol and atovaquone show synergistic interaction. Accordingly, atovaquone-mediated disruption of mitochondrial membrane potential reduces the import and recruitment of both subunits of PfTopoVI to mitochondrial DNA (mtDNA) in a dose-dependent manner. The structural differences between PfTopoVIB and human TopoVIB-like protein could be exploited for development of a novel antimalarial agent. IMPORTANCE This study demonstrates a likely role of topoisomerase VI in the mitochondrial genome segregation of Plasmodium falciparum during endoreduplication. We show that PfTopoVIB and PfSpo11 remain associated and form the functional holoenzyme within the parasite. The spatiotemporal expression of both subunits of PfTopoVI correlates well with their recruitment to the mitochondrial DNA at the late schizont stage of the parasite. Additionally, the synergistic interaction between PfTopoVI inhibitor and the disruptor of mitochondrial membrane potential, atovaquone, supports that topoisomerase VI is the mitochondrial topoisomerase of the malaria parasite. We propose that topoisomerase VI may act as a novel target against malaria. |
| first_indexed | 2024-03-13T05:20:34Z |
| format | Article |
| id | doaj.art-d42e1672c0364164b309bdd8ba2bc3e1 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-03-13T05:20:34Z |
| publishDate | 2023-06-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-d42e1672c0364164b309bdd8ba2bc3e12023-06-15T13:18:33ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-06-0111310.1128/spectrum.04980-22Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA SegregationPriyanka Singh0Wahida Tabassum1Nupur Fangaria2Sandeep Dey3Siladitya Padhi4Mrinal K. Bhattacharyya5Kota Arun Kumar6Arijit Roy7Sunanda Bhattacharyya8Department of Biotechnology and Bioinformatics, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaDepartment of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaDepartment of Biotechnology and Bioinformatics, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaDepartment of Animal Biology, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaTCS Research-Hyderabad (Life Sciences Division), Tata Consultancy Services Limited, Hyderabad, IndiaDepartment of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaDepartment of Animal Biology, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaTCS Research-Hyderabad (Life Sciences Division), Tata Consultancy Services Limited, Hyderabad, IndiaDepartment of Biotechnology and Bioinformatics, School of Life Sciences, University of Hyderabad, Hyderabad, IndiaABSTRACT The human malaria parasite undergoes a noncanonical cell division, namely, endoreduplication, where several rounds of nuclear, mitochondrial, and apicoplast replication occur without cytoplasmic division. Despite its importance in Plasmodium biology, the topoisomerases essential for decatenation of replicated chromosome during endoreduplication remain elusive. We hypothesize that the topoisomerase VI complex, containing Plasmodium falciparum topiosomerase VIB (PfTopoVIB) and catalytic P. falciparum Spo11 (PfSpo11), might be involved in the segregation of the Plasmodium mitochondrial genome. Here, we demonstrate that the putative PfSpo11 is the functional ortholog of yeast Spo11 that can complement the sporulation defects of the yeast Δspo11 strain, and the catalytic mutant Pfspo11Y65F cannot complement such defects. PfTopoVIB and PfSpo11 display a distinct expression pattern compared to the other type II topoisomerases of Plasmodium and are induced specifically at the late schizont stage of the parasite, when the mitochondrial genome segregation occurs. Furthermore, PfTopoVIB and PfSpo11 are physically associated with each other at the late schizont stage, and both subunits are localized in the mitochondria. Using PfTopoVIB- and PfSpo11-specific antibodies, we immunoprecipitated the chromatin of tightly synchronous early, mid-, and late schizont stage-specific parasites and found that both the subunits are associated with the mitochondrial genome during the late schizont stage of the parasite. Furthermore, PfTopoVIB inhibitor radicicol and atovaquone show synergistic interaction. Accordingly, atovaquone-mediated disruption of mitochondrial membrane potential reduces the import and recruitment of both subunits of PfTopoVI to mitochondrial DNA (mtDNA) in a dose-dependent manner. The structural differences between PfTopoVIB and human TopoVIB-like protein could be exploited for development of a novel antimalarial agent. IMPORTANCE This study demonstrates a likely role of topoisomerase VI in the mitochondrial genome segregation of Plasmodium falciparum during endoreduplication. We show that PfTopoVIB and PfSpo11 remain associated and form the functional holoenzyme within the parasite. The spatiotemporal expression of both subunits of PfTopoVI correlates well with their recruitment to the mitochondrial DNA at the late schizont stage of the parasite. Additionally, the synergistic interaction between PfTopoVI inhibitor and the disruptor of mitochondrial membrane potential, atovaquone, supports that topoisomerase VI is the mitochondrial topoisomerase of the malaria parasite. We propose that topoisomerase VI may act as a novel target against malaria.https://journals.asm.org/doi/10.1128/spectrum.04980-22Plasmodium mitochondriaPlasmodium topoisomerase VIapicomplexan topoisomerase |
| spellingShingle | Priyanka Singh Wahida Tabassum Nupur Fangaria Sandeep Dey Siladitya Padhi Mrinal K. Bhattacharyya Kota Arun Kumar Arijit Roy Sunanda Bhattacharyya Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation Microbiology Spectrum Plasmodium mitochondria Plasmodium topoisomerase VI apicomplexan topoisomerase |
| title | Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation |
| title_full | Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation |
| title_fullStr | Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation |
| title_full_unstemmed | Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation |
| title_short | Plasmodium Topoisomerase VIB and Spo11 Constitute Functional Type IIB Topoisomerase in Malaria Parasite: Its Possible Role in Mitochondrial DNA Segregation |
| title_sort | plasmodium topoisomerase vib and spo11 constitute functional type iib topoisomerase in malaria parasite its possible role in mitochondrial dna segregation |
| topic | Plasmodium mitochondria Plasmodium topoisomerase VI apicomplexan topoisomerase |
| url | https://journals.asm.org/doi/10.1128/spectrum.04980-22 |
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