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Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes

BACKGROUND: Malaria is a major global parasitic disease caused by species of the genus Plasmodium. Zygotes of Plasmodium spp. undergo meiosis and develop into tetraploid ookinetes, which differentiate into oocysts that undergo sporogony. Homologous recombination (HR) occurs during meiosis and introd...

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Autores principales: Yoshikawa, Yasunaga, Kimura, Shunta, Soga, Akira, Sugiyama, Makoto, Ueno, Aki, Kondo, Hiroki, Zhu, Zida, Ochiai, Kazuhiko, Nakayama, Kazuhiko, Hakozaki, Jun, Kusakisako, Kodai, Haraguchi, Asako, Kitano, Taisuke, Orino, Koichi, Fukumoto, Shinya, Ikadai, Hiromi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9270840/
https://www.ncbi.nlm.nih.gov/pubmed/35804459
http://dx.doi.org/10.1186/s13071-022-05357-w
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author Yoshikawa, Yasunaga
Kimura, Shunta
Soga, Akira
Sugiyama, Makoto
Ueno, Aki
Kondo, Hiroki
Zhu, Zida
Ochiai, Kazuhiko
Nakayama, Kazuhiko
Hakozaki, Jun
Kusakisako, Kodai
Haraguchi, Asako
Kitano, Taisuke
Orino, Koichi
Fukumoto, Shinya
Ikadai, Hiromi
author_facet Yoshikawa, Yasunaga
Kimura, Shunta
Soga, Akira
Sugiyama, Makoto
Ueno, Aki
Kondo, Hiroki
Zhu, Zida
Ochiai, Kazuhiko
Nakayama, Kazuhiko
Hakozaki, Jun
Kusakisako, Kodai
Haraguchi, Asako
Kitano, Taisuke
Orino, Koichi
Fukumoto, Shinya
Ikadai, Hiromi
author_sort Yoshikawa, Yasunaga
collection PubMed
description BACKGROUND: Malaria is a major global parasitic disease caused by species of the genus Plasmodium. Zygotes of Plasmodium spp. undergo meiosis and develop into tetraploid ookinetes, which differentiate into oocysts that undergo sporogony. Homologous recombination (HR) occurs during meiosis and introduces genetic variation. However, the mechanisms of HR in Plasmodium are unclear. In humans, the recombinases DNA repair protein Rad51 homolog 1 (Rad51) and DNA meiotic recombinase 1 (Dmc1) are required for HR and are regulated by breast cancer susceptibility protein 2 (BRCA2). Most eukaryotes harbor BRCA2 homologs. Nevertheless, these have not been reported for Plasmodium. METHODS: A Brca2 candidate was salvaged from a database to identify Brca2 homologs in Plasmodium. To confirm that the candidate protein was Brca2, interaction activity between Plasmodium berghei (Pb) Brca2 (PbBrca2) and Rad51 (PbRad51) was investigated using a mammalian two-hybrid assay. To elucidate the functions of PbBrca2, PbBrca2 was knocked out and parasite proliferation and differentiation were assessed in mice and mosquitoes. Transmission electron microscopy was used to identify sporogony. RESULTS: The candidate protein was conserved among Plasmodium species, and it was indicated that it harbors critical BRCA2 domains including BRC repeats, tower, and oligonucleotide/oligosaccharide-binding-fold domains. The P. berghei BRC repeats interacted with PbRad51. Hence, the candidate was considered a Brca2 homolog. PbBrca2 knockout parasites were associated with reduced parasitemia with increased ring stage and decreased trophozoite stage counts, gametocytemia, female gametocyte ratio, oocyst number, and ookinete development in both mice and mosquitoes. Nevertheless, the morphology of the blood stages in mice and the ookinete stage was comparable to those of the wild type parasites. Transmission electron microscopy results showed that sporogony never progressed in Brca2-knockout parasites. CONCLUSIONS: Brca2 is implicated in nearly all Plasmodium life cycle stages, and especially in sporogony. PbBrca2 contributes to HR during meiosis. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-022-05357-w.
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spelling pubmed-92708402022-07-10 Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes Yoshikawa, Yasunaga Kimura, Shunta Soga, Akira Sugiyama, Makoto Ueno, Aki Kondo, Hiroki Zhu, Zida Ochiai, Kazuhiko Nakayama, Kazuhiko Hakozaki, Jun Kusakisako, Kodai Haraguchi, Asako Kitano, Taisuke Orino, Koichi Fukumoto, Shinya Ikadai, Hiromi Parasit Vectors Research BACKGROUND: Malaria is a major global parasitic disease caused by species of the genus Plasmodium. Zygotes of Plasmodium spp. undergo meiosis and develop into tetraploid ookinetes, which differentiate into oocysts that undergo sporogony. Homologous recombination (HR) occurs during meiosis and introduces genetic variation. However, the mechanisms of HR in Plasmodium are unclear. In humans, the recombinases DNA repair protein Rad51 homolog 1 (Rad51) and DNA meiotic recombinase 1 (Dmc1) are required for HR and are regulated by breast cancer susceptibility protein 2 (BRCA2). Most eukaryotes harbor BRCA2 homologs. Nevertheless, these have not been reported for Plasmodium. METHODS: A Brca2 candidate was salvaged from a database to identify Brca2 homologs in Plasmodium. To confirm that the candidate protein was Brca2, interaction activity between Plasmodium berghei (Pb) Brca2 (PbBrca2) and Rad51 (PbRad51) was investigated using a mammalian two-hybrid assay. To elucidate the functions of PbBrca2, PbBrca2 was knocked out and parasite proliferation and differentiation were assessed in mice and mosquitoes. Transmission electron microscopy was used to identify sporogony. RESULTS: The candidate protein was conserved among Plasmodium species, and it was indicated that it harbors critical BRCA2 domains including BRC repeats, tower, and oligonucleotide/oligosaccharide-binding-fold domains. The P. berghei BRC repeats interacted with PbRad51. Hence, the candidate was considered a Brca2 homolog. PbBrca2 knockout parasites were associated with reduced parasitemia with increased ring stage and decreased trophozoite stage counts, gametocytemia, female gametocyte ratio, oocyst number, and ookinete development in both mice and mosquitoes. Nevertheless, the morphology of the blood stages in mice and the ookinete stage was comparable to those of the wild type parasites. Transmission electron microscopy results showed that sporogony never progressed in Brca2-knockout parasites. CONCLUSIONS: Brca2 is implicated in nearly all Plasmodium life cycle stages, and especially in sporogony. PbBrca2 contributes to HR during meiosis. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-022-05357-w. BioMed Central 2022-07-08 /pmc/articles/PMC9270840/ /pubmed/35804459 http://dx.doi.org/10.1186/s13071-022-05357-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Yoshikawa, Yasunaga
Kimura, Shunta
Soga, Akira
Sugiyama, Makoto
Ueno, Aki
Kondo, Hiroki
Zhu, Zida
Ochiai, Kazuhiko
Nakayama, Kazuhiko
Hakozaki, Jun
Kusakisako, Kodai
Haraguchi, Asako
Kitano, Taisuke
Orino, Koichi
Fukumoto, Shinya
Ikadai, Hiromi
Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes
title Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes
title_full Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes
title_fullStr Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes
title_full_unstemmed Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes
title_short Plasmodium berghei Brca2 is required for normal development and differentiation in mice and mosquitoes
title_sort plasmodium berghei brca2 is required for normal development and differentiation in mice and mosquitoes
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9270840/
https://www.ncbi.nlm.nih.gov/pubmed/35804459
http://dx.doi.org/10.1186/s13071-022-05357-w
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