Blocking palmitoylation of<i>Toxoplasma gondii</i>myosin light chain 1 disrupts glideosome composition but has little impact on parasite motility

Rompikuntal, Pramod Kumar; Foe, Ian T.; Deng, Bin; Bogyo, Matthew; Ward, Gary E.

doi:10.1101/2020.08.13.250399

Cited by 3 publications

(3 citation statements)

References 71 publications

(122 reference statements)

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“…This would suggest that palmitoylation only partially contributes to concavin function and could hint towards the presence of other protein-protein interaction that are essential for concavin functionality and sporozoite shape maintenance. This finding of partial impact of palmitoylation is reminiscent of the recent work on palmitoylation of the myosin light chain in Toxoplasma gondii (Rompikuntal et al, 2020), where disruption of palmitoylation led to the partial disassembly of the gliding motor complex but had little impact on motility itself. Considering the EM images showing large invaginations of the IMC away from the plasma membrane in concavin(-) sporozoites, we postulate that concavin might contribute towards riveting the plasma membrane to the subtending IMC, which are a constant distance of 25 nm apart.…”

Section: Discussionsupporting

confidence: 61%

Malaria transmission relies on concavin-mediated maintenance ofPlasmodiumsporozoite cell shape

Kehrer

Formaglio

Muthinja

et al. 2021

Preprint

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During transmission of malaria-causing parasites from mosquitoes to mammals, Plasmodium sporozoites migrate rapidly in the skin to search for a blood vessel. The high migratory speed and narrow passages taken by the parasites suggest considerable strain on the sporozoites to maintain their shape. Here we report on a newly identified protein, concavin, that is important for maintenance of the sporozoite shape inside salivary glands of mosquitoes and during migration in the skin. Concavin-GFP localized at the cytoplasmic periphery of sporozoites and concavin(−) sporozoites progressively rounded up upon entry of salivary glands. These rounded concavin(−) sporozoites failed to pass through the narrow salivary ducts and were hence rarely ejected by mosquitoes. However, normally shaped concavin(−) sporozoites could be transmitted and migrated in the skin or skin like environments. Strikingly, motile concavin(−) sporozoites could disintegrate while migrating through narrow strictures in the skin leading to parasite arrest or death and decreased transmission efficiency. We suggest that concavin contributes to cell shape maintenance by riveting the plasma membrane to the subtending inner membrane complex.SIGNIFICANCEMalaria parasites are transmitted by Anopheles mosquitoes and rely on rapid migration for establishing an infection. We identified and characterized a protein, named concavin, essential for maintaining the shape of the sporozoite. Concavin is a membrane associated protein facing the cytoplasm suggesting that it contributes to riveting the plasma membrane to the subtending inner membrane complex. Sporozoites lacking concavin can round up in the salivary glands, are less well transmitted to mice and disintegrate while migrating in the skin. Hence, concavin is essential for parasite transmission and infectivity.Highlights- A membrane associated protein is essential for Plasmodium shape maintenance-Migrating parasites disintegrate in the absence of concavin-First protein essential for cellular integrity of Plasmodium sporozoites-Thickened and deformed Plasmodium sporozoites fail to be transmitted by mosquitoes

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Section: Discussionsupporting

confidence: 61%

Malaria transmission relies on concavin-mediated maintenance ofPlasmodiumsporozoite cell shape

Kehrer

Formaglio

Muthinja

et al. 2021

Preprint

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“…The reactive cysteine identified in Tg ISPH contributes to an iron-sulfur cluster that is required for enzyme catalysis, and therefore any substitutions at this site are expected to be deleterious. Tg MLC1 is part of the glideosome complex required for parasite motility and host-cell invasion 31 and contains two N-terminal reactive cysteines that are known to be S -acylated 32,33 yet are non-essential for Tg MLC1 function 34 . We therefore hypothesized that substitutions at this site would not affect parasite fitness.…”

Section: Main Textmentioning

confidence: 99%

Prioritization of antimicrobial targets by CRISPR-based oligo recombineering

Benns

Storch

Falco

et al. 2021

Preprint

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Nucleophilic amino acids are important in covalent drug development yet underutilized as antimicrobial targets. Over recent years, several chemoproteomic technologies have been developed to mine chemically-accessible residues via their intrinsic reactivity toward electrophilic probes. However, these approaches cannot discern which reactive sites contribute to protein function and should therefore be prioritized for drug discovery. To address this, we have developed a CRISPR-based Oligo Recombineering (CORe) platform to systematically prioritize reactive amino acids according to their contribution to protein function. Our approach directly couples protein sequence and function with biological fitness. Here, we profile the reactivity of >1,000 cysteines on ~700 proteins in the eukaryotic pathogen Toxoplasma gondii and prioritize functional sites using CORe. We competitively compared the fitness effect of 370 codon switches at 74 cysteines and identify functional sites in a diverse range of proteins. In our proof of concept, CORe performed >800 times faster than a standard genetic workflow. Reactive cysteines decorating the ribosome were found to be critical for parasite growth, with subsequent target-based screening validating the apicomplexan translation machinery as a target for covalent ligand development. CORe is system-agnostic, and supports expedient identification, functional prioritization, and rational targeting of reactive sites in a wide range of organisms and diseases.

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“…Likewise, the absence of GAP45 in gregarines and Cryptosporidium maybe compensated by other GAP-like proteins or even not be a problem at all; indeed, it has been proposed that a motor architecture could be organized in a much looser manner, in which actin-myosin motors push in a general backward direction, but without necessarily being guided by GAP proteins (Tardieux and Baum, 2016). Furthermore, while TgMLC1 binding to TgGAP45 is considered a key component of the parasite's force transduction mechanism, it has recently been shown that loss of TgMLC1 binding to TgGAP45 has little effect on their ability to initiate or maintain movement 102 , questioning again the real role of GAP45 and suggesting our comprehension of the glideosome's proteins' intrication is still incomplete.…”

Section: The Gregarine Glideosome(s)mentioning

confidence: 99%

Marine gregarine genomes reveal the breadth of apicomplexan diversity and provide new insights on gliding motility

Boisard¹,

Duvernois-Berthet

Duval

et al. 2021

Preprint

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Apicomplexans, parasite protists of a very wide diversity of metazoan hosts, are mostly known from species infecting human. Absence or limited data for basal lineages prevents a comprehensive view of evolutionary history and adaptive capacities of Apicomplexa. Here, we characterized the genome of the marine eugregarine Porospora gigantea, remarkable for the gigantic size of its vegetative feeding forms (trophozoites) and their speed of gliding movement, the fastest so far recorded for an Apicomplexa. Not a single but two highly related genomes named A and B were assembled. Highly syntenic, of similar size (9 Mb) and coding capacities (~5300 genes), they display a 10.8% divergence at nucleotide level corresponding to 16-38 My divergent time. Orthogroups analyses across 25 (proto)Apicomplexa including Gregarina niphandrodes showed that A and B are highly divergent from all other known apicomplexan species, revealing an unexpected diversity. These two related species branch in phylogenetic studies at the base of Cephaloidophoroidea, forming a new family in these crustacean gregarines. Gliding proteins data mining found a strong conservation of actin-related proteins, as well as of regulatory factors, within apicomplexan. In contrast, the conservation of core glideosome proteins and adhesion proteins appears to be highly variable among apicomplexan lineages, especially in gregarines. These results confirm the importance of studying gregarines to widen our biological and evolutionary view of apicomplexan parasites, to better apprehend species diversity and revise our understanding of the molecular bases of some key functions such as observed for the glideosome.

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Blocking palmitoylation ofToxoplasma gondiimyosin light chain 1 disrupts glideosome composition but has little impact on parasite motility

Cited by 3 publications

References 71 publications

Malaria transmission relies on concavin-mediated maintenance ofPlasmodiumsporozoite cell shape

Malaria transmission relies on concavin-mediated maintenance ofPlasmodiumsporozoite cell shape

Prioritization of antimicrobial targets by CRISPR-based oligo recombineering

Marine gregarine genomes reveal the breadth of apicomplexan diversity and provide new insights on gliding motility

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