Composition and organization of kinetochores show plasticity in apicomplexan chromosome segregation

Brusini, Lorenzo; Pacheco, Nicolas Dos Santos; Tromer, Eelco C.; Soldati‐Favre, Dominique; Brochet, Mathieu

doi:10.1083/jcb.202111084

Cited by 24 publications

(36 citation statements)

References 95 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this study using PyEB1 as a spindle marker, we observed that the kinetochores were localized to the lateral side of hemispindle MTs throughout three rounds of endomitosis. Consistent with it, several recent studies had also revealed similar MT lateral positioning of kinetochores in the erythrocytic schizogony and male gametogenesis [9, 63]. Interestingly, in the human cells with artificially induced monopolar spindle, which structurally mimics the hemispindle of the Plasmodium , lateral attachment of kinetochores with MT were also detected as an intermediate status during the lateral to end-on attachment conversion [64].…”

Section: Discussionsupporting

confidence: 54%

EB1 decoration of microtubule lattice facilitates spindle-kinetochore lateral attachment inPlasmodiummale gametogenesis

Yang

Cai

Huang

et al. 2023

Preprint

View full text Add to dashboard Cite

Faithful chromosome segregation of 8 duplicated haploid genomes into 8 daughter male gametes is essential for male gametogenesis and mosquito transmission of Plasmodium. Plasmodium evolves the endomitosis for this multinucleated cell division. However, the mechanism underlying the spindle-kinetochore attachment remains elusive. End-binding proteins (EBs) are the conserved microtubule (MT) plus-end binding proteins and play an important role in regulating MT plus-end dynamics. Here we report that Plasmodium EB1 is a unique orthologue distinct from the canonical eukaryotic EB1. Both in vitro and in vivo assays revealed that Plasmodium EB1 lost MT plus-end tracking but gained MT-lattice affinity. This MT-binding feature of EB1 is contributed by both the CH domain and the linker region. EB1-deficient parasites produce male gametocytes that develop to the anucleated male gametes, leading to defective mosquito transmission of parasite. EB1 is localized at the nucleoplasm of male gametocytes. Upon gametogenesis, EB1 decorates the full-length of spindle MTs and regulates spindle structure. The kinetochores attach to spindle MTs laterally throughout three rounds of endomitosis and this attachment is EB1-dependent. Consequently, impaired spindle-kinetochore attachment was observed in EB1-deficient parasites. These results indicate that a parasite-specific EB1 with MT-lattice affinity has evolved to fulfill the spindle-kinetochore lateral attachment in male gametogenesis.

show abstract

Section: Discussionsupporting

confidence: 54%

EB1 decoration of microtubule lattice facilitates spindle-kinetochore lateral attachment inPlasmodiummale gametogenesis

Yang

Cai

Huang

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Generally, few unique peptides were identified for each protein except for ARK2 itself, suggesting that their binding to ARK2 may be transient or that much of ARK2 is not bound to other proteins at this stage. The interacting proteins identified include the spindle MT-associated proteins kinesin-8X (PBANKA_0805900), myosin K (PBANKA_0908500), the MT plus-end tracker EB1 (PBANKA_0405600), a variety of kinetochore proteins such as members of the NDC80 complex (Zeeshan et al, 2020b) and the recently discovered highly divergent Apicomplexan Kinetochore proteins (AKiT) AKiT1-6, STU2 (PBANKA_1337500), Mad1 (PBANKA_0612300) (Brusini et al, 2022), and a single peptide for the CPC subunit INCENP2 (PBANKA_1343200) S3). Interestingly, the strongest evidence for an ARK2 interaction was obtained for the nuclear formin-like protein MISFIT, a key regulator of ookinete-oocyst transition (Bushell et al, 2009), and a putative regulator of actin filament dynamics.…”

Section: Ark2 Interacts With Microtubule-binding Proteins Near the Sp...mentioning

confidence: 99%

Plasmodium ARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation and chromosome segregation of sexual transmission stages

Zeeshan¹,

Rea

Abel

et al. 2023

Preprint

View full text Add to dashboard Cite

Mechanisms of cell division are remarkably diverse, suggesting the underlying molecular networks among eukaryotes differ extensively. The Aurora family of kinases orchestrates the process of chromosome segregation and cytokinesis during cell division through precise spatiotemporal regulation of their catalytic activities by distinct scaffolds. Plasmodium spp., the causative agents of malaria, are unicellular eukaryotes that have three divergent aurora-related kinases (ARKs) and lack most canonical scaffolds/activators. The parasite uses unconventional modes of chromosome segregation during endomitosis and meiosis in sexual transmission stages within mosquito host. This includes a rapid threefold genome replication from 1N to 8N with successive cycles of closed mitosis, spindle formation and chromosome segregation within eight minutes (termed male gametogony). Kinome studies had previously suggested likely essential functions for all three Plasmodium ARKs during asexual mitotic cycles; however, little is known about their location, function, or their scaffolding molecules during unconventional sexual proliferative stages. Using a combination of super-resolution microscopy, mass spectrometry, omics and live-cell fluorescence imaging, we set out to investigate the contribution of the atypical Aurora paralog ARK2 to proliferative sexual stages using rodent malaria model Plasmodium berghei. We find that ARK2 primarily localises to the spindle apparatus associated with kinetochores during both mitosis and meiosis. Interactomics and co-localisation studies reveal a unique ARK2 scaffold at the spindle including the microtubule plus end-binding protein EB1 and lacking some other conserved molecules. Gene function studies indicate complementary functions of ARK2 and EB1 in driving endomitotic divisions and thereby parasite transmission. Our discovery of a novel Aurora spindle scaffold underlines the emerging flexibility of molecular networks to rewire and drive unconventional mechanisms of chromosome segregation in the malaria parasite Plasmodium.

show abstract

“…(PBANKA_1337500), Mad1 (PBANKA_0612300) (Brusini et al, 2022), and a single peptide for the CPC subunit INCENP2 (PBANKA_1343200) (Fig 5B, Table S3).…”

Section: Ark2 Interacts With Microtubule-binding Proteins Near the Sp...mentioning

confidence: 99%

PlasmodiumARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation and chromosome segregation of sexual transmission stages

Zeeshan

Rea

Abel

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Mechanisms of cell division are remarkably diverse, suggesting the underlying molecular networks among eukaryotes differ extensively. The Aurora family of kinases orchestrates the process of chromosome segregation and cytokinesis during cell division through precise spatiotemporal regulation of their catalytic activities by distinct scaffolds. Plasmodium spp., the causative agents of malaria, are unicellular eukaryotes that have three divergent aurora-related kinases (ARKs) and lack most canonical scaffolds/activators. The parasite uses unconventional modes of chromosome segregation during endomitosis and meiosis in sexual transmission stages within mosquito host. This includes a rapid threefold genome replication from 1N to 8N with successive cycles of closed mitosis, spindle formation and chromosome segregation within eight minutes (termed male gametogony). Kinome studies had previously suggested likely essential functions for all three Plasmodium ARKs during asexual mitotic cycles; however, little is known about their location, function, or their scaffolding molecules during unconventional sexual proliferative stages. Using a combination of super-resolution microscopy, mass spectrometry, and live-cell fluorescence imaging, we set out to investigate the role of the atypical Aurora paralog ARK2 to proliferative sexual stages using rodent malaria model Plasmodium berghei. We find that ARK2 primarily localises to the spindle apparatus in the vicinity of kinetochores during both mitosis and meiosis. Interactomics and co-localisation studies reveal a unique ARK2 scaffold at the spindle including the microtubule plus end-binding protein EB1, lacking conserved Aurora scaffold proteins. Gene function studies indicate complementary functions of ARK2 and EB1 in driving endomitotic divisions and thereby parasite transmission. Our discovery of a novel Aurora kinase spindle scaffold underlines the emerging flexibility of molecular networks to rewire and drive unconventional mechanisms of chromosome segregation in the malaria parasite Plasmodium.

show abstract

Composition and organization of kinetochores show plasticity in apicomplexan chromosome segregation

Cited by 24 publications

References 95 publications

EB1 decoration of microtubule lattice facilitates spindle-kinetochore lateral attachment inPlasmodiummale gametogenesis

EB1 decoration of microtubule lattice facilitates spindle-kinetochore lateral attachment inPlasmodiummale gametogenesis

Plasmodium ARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation and chromosome segregation of sexual transmission stages

PlasmodiumARK2-EB1 axis drives the unconventional spindle dynamics, scaffold formation and chromosome segregation of sexual transmission stages

Contact Info

Product

Resources

About