Generation of Plasmodium yoelii malaria parasite for conditional degradation of proteins

Liu, Chuanyuan; Yang, Zhenke; Cai, Mengya; Shi, Yang; Cui, Huiting; Yuan, Jing

doi:10.1016/j.molbiopara.2020.111346

Cited by 10 publications

(10 citation statements)

References 30 publications

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“…This observation is in agreement with the recent global knockout screening results in Plasmodium falciparum and Plasmodium berghei (44,45). Therefore, we applied an auxin-inducible degron (AID)-based protein degradation system we recently adapted in P. yoelii (46), which allows rapid depletion of a target protein fused to an AID motif with the aid of auxin/indole-3-acetic acid (IAA) (47). The C terminus of the endogenous cdc50c locus was tagged with the sequence encoding the AID::6HA in the soap-Tir1 strain (fig.…”

Section: Cdc50c Depletion In Ookinetes Phenocopies Atp7 Deficiency During Mosquito Transmissionsupporting

confidence: 86%

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A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission

et al. 2021

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Mosquito midgut epithelium traversal is essential for malaria parasite transmission. Phospholipid flippases are eukaryotic type 4 P-type adenosine triphosphatases (P4-ATPases), which, in association with CDC50, translocate phospholipids across the membrane lipid bilayers. In this study, we investigated the function of a putative P4-ATPase, ATP7, from the rodent malaria parasite Plasmodium yoelii. Disruption of ATP7 blocks the parasite infection of mosquitoes. ATP7 is localized on the ookinete plasma membrane. While ATP7-depleted ookinetes are capable of invading the midgut, they are eliminated within the epithelial cells by a process independent from the mosquito complement-like immunity. ATP7 colocalizes and interacts with the flippase cofactor CDC50C. Depletion of CDC50C phenocopies ATP7 deficiency. ATP7-depleted ookinetes fail to uptake phosphatidylcholine across the plasma membrane. Ookinete microinjection into the mosquito hemocoel reverses the ATP7 deficiency phenotype. Our study identifies Plasmodium flippase as a mechanism of parasite survival in the midgut epithelium that is required for mosquito transmission.

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Section: Cdc50c Depletion In Ookinetes Phenocopies Atp7 Deficiency During Mosquito Transmissionsupporting

confidence: 86%

“…The C terminus of the endogenous cdc50c locus was tagged with the sequence encoding the AID::6HA in the soap-Tir1 strain (fig. S1) (46). The resulting cdc50c::aid parasite exhibited the expected cell periphery localization of CDC50C in ookinetes (Fig.…”

Section: Cdc50c Depletion In Ookinetes Phenocopies Atp7 Deficiency During Mosquito Transmissionmentioning

confidence: 78%

A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission

et al. 2021

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“…DHHC2 has been suggested to play an essential role in the asexual blood stage development of the parasites since no viable mutant clone was obtained using either the conventional or Cas9-based knockout strategies in the P. yoelii and P. berghei [17, 55]. To explore the functions of DHHC2, we applied an Auxin-inducible degron (AID)-based protein degradation system in the P. yoelii transgenic strain Tir1 [56], which allows depletion of the target protein fused to a miniAID (mAID) motif with the aid of the plant hormone auxin (Indole-3-acetic acid, IAA). The N-terminus of the endogenous dhhc2 locus was tagged with the sequence encoding mAID::2HA in the Tir1 strain, generating the mAID::dhhc2 clone (Fig S6A).…”

Section: Resultsmentioning

confidence: 99%

Inner membrane complex proteomics reveals a palmitoylation cascade regulating intraerythrocytic development of malaria parasite

Qian

Wang

Zhong

et al. 2022

Preprint

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Malaria is caused by infection of the erythrocytes by the parasites Plasmodium. Inside the erythrocytes, the parasites multiply via schizogony, an unconventional cell division mode. The Inner Membrane Complex (IMC), an organelle located beneath the parasite plasma membrane, serving as the platform for protein anchorage, is essential for schizogony. So far, complete repertoire of IMC proteins and their localization determinants remain unclear. Here we used biotin ligase (TurboID)-based proximity labelling to compile the proteome of the schizont IMC of rodent malaria parasite Plasmodium yoelii. In total, 300 TurboID-interacting proteins were identified. 19 of the 22 selected candidates were confirmed to localize in the IMC, indicating good reliability. In light of the existing palmitome of Plasmodium falciparum, 83 proteins of the P. yoelii IMC proteome are potentially palmitoylated. We further identified DHHC2 as the major resident palmitoyl-acyl-transferase of the IMC. Depletion of DHHC2 led to defective schizont segmentation and growth arrest both in vitro and in vivo. DHHC2 was found to palmitoylate two critical IMC proteins CDPK1 and GAP45 for their IMC localization. In summary, this study reports an inventory of new IMC proteins and demonstrates a central role of DHHC2 in governing IMC localization of proteins during the schizont development.

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“…Although not yet adapted for high-throughput assays, several conditional systems have been developed in P. berghei and P. yoelii that may permit functional screening at all life cycle stages regardless of essentiality. The AID-system for protein degradation ( Philip and Waters, 2015 ; Liu et al., 2021 ) could be integrated into the endogenous tagging pipeline of the recombineering platform ( Pfander et al., 2011 ) to enable simultaneous localisation and control of protein levels by addition of IAA. While knockdown of AID tagged proteins has been achieved in the peritoneum of mice ( Brown and Sibley, 2018 ), more biologically relevant knockdowns have yet to be demonstrated, possibly due to the high dose of IAA required to induce a knockdown and its toxicity for rodents in in vivo studies ( Yesbolatova et al., 2020 ).…”

Section: Functional Screeningmentioning

confidence: 99%

Paving the Way: Contributions of Big Data to Apicomplexan and Kinetoplastid Research

Kent

Briggs²,

Colon³

et al. 2022

Front. Cell. Infect. Microbiol.

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In the age of big data an important question is how to ensure we make the most out of the resources we generate. In this review, we discuss the major methods used in Apicomplexan and Kinetoplastid research to produce big datasets and advance our understanding of Plasmodium, Toxoplasma, Cryptosporidium, Trypanosoma and Leishmania biology. We debate the benefits and limitations of the current technologies, and propose future advancements that may be key to improving our use of these techniques. Finally, we consider the difficulties the field faces when trying to make the most of the abundance of data that has already been, and will continue to be, generated.

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Generation of Plasmodium yoelii malaria parasite for conditional degradation of proteins

Cited by 10 publications

References 30 publications

A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission

A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission

Inner membrane complex proteomics reveals a palmitoylation cascade regulating intraerythrocytic development of malaria parasite

Paving the Way: Contributions of Big Data to Apicomplexan and Kinetoplastid Research

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