bims-toxgon 2023-12-10 papers

bims-toxgon

Biomed News

on Toxoplasma gondii metabolism

Issue of 2023–12–10
sixteen papers selected by
Lakesh Kumar, BITS Pilani

A Toxoplasma gondii putative amino acid transporter localizes to the plant-like vacuolar compartment and controls parasite extracellular survival and stage differentiation.
Toxoplasma rhoptry proteins that affect encephalitis outcome.
Evaluation of topotecan and 10-hydroxycamptothecin on Toxoplasma gondii: Implications on baseline DNA damage and repair efficiency.
Proteomics analysis of Toxoplasma gondii merozoites reveals regulatory proteins involved in sexual reproduction.
The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques.
Toxoplasma gondii Infection in Pregnancy - Recommendations of the Working Group on Obstetrics and Prenatal Medicine (AGG - Section on Maternal Disorders).
Plasmodium yoelii iron transporter PyDMT1 interacts with host ferritin and is required in full activity for malarial pathogenesis.
Plasmodium falciparum utilizes pyrophosphate to fuel an essential proton pump in the ring stage and the transition to trophozoite stage.
Whole genome enrichment approach for genomic surveillance of Toxoplasma gondii.
Effects of the G-quadruplex-binding drugs quarfloxin and CX-5461 on the malaria parasite Plasmodium falciparum.
Editorial: Lipids in host and protozoan parasite interaction.
Histone deacetylase 6: at the interface of cancer and neurodegeneration.
mTOR takes charge: Relaying uncharged tRNA levels by mTOR ubiquitination.
Drug addiction and treatment: An epigenetic perspective.
Lysine acetylation regulates the AT-rich DNA possession ability of H-NS.
Learning sequence, structure, and function representations of proteins with language models.

mSphere. 2023 Dec 05. e0059723

A Toxoplasma gondii putative amino acid transporter localizes to the plant-like vacuolar compartment and controls parasite extracellular survival and stage differentiation.

Federica Piro, Silvia Masci, Geetha Kannan, Riccardo Focaia, Tracey L Schultz, Pariyamon Thaprawat, Vern B Carruthers, Manlio Di Cristina.

   IMPORTANCE: Toxoplasma gondii is a highly successful parasite infecting a broad range of warm-blooded organisms, including about one-third of all humans. Although Toxoplasma infections rarely result in symptomatic disease in individuals with a healthy immune system, the incredibly high number of persons infected, along with the risk of severe infection in immunocompromised patients and the potential link of chronic infection to mental disorders, makes this infection a significant public health concern. As a result, there is a pressing need for new treatment approaches that are both effective and well tolerated. The limitations in understanding how Toxoplasma gondii manages its metabolism to adapt to changing environments and triggers its transformation into bradyzoites have hindered the discovery of vulnerabilities in its metabolic pathways or nutrient acquisition mechanisms to identify new therapeutic targets. In this work, we have shown that the lysosome-like organelle plant-like vacuolar compartment (PLVAC), acting through the putative arginine transporter TgAAT1, plays a pivotal role in regulating the parasite's extracellular survival and differentiation into bradyzoites.

Keywords:  SLC38A9; TORC complex; Toxoplasma gondii; amino acid transporters; arginine; bradyzoite differentiation; extracellular survival

DOI:  https://doi.org/10.1128/msphere.00597-23
Cell Death Discov. 2023 Dec 04. 9(1): 439

Toxoplasma rhoptry proteins that affect encephalitis outcome.

Xinlei Wang, Lai Qu, Jie Chen, Yufen Jin, Kaisong Hu, Zhengjie Zhou, Jiaqi Zhang, Yiming An, Jingtong Zheng.

Toxoplasma gondii, a widespread obligate intracellular parasite, can infect almost all warm-blooded animals, including humans. The cellular barrier of the central nervous system (CNS) is generally able to protect the brain parenchyma from infectious damage. However, T. gondii typically causes latent brain infections in humans and other vertebrates. Here, we discuss how T. gondii rhoptry proteins (ROPs) affect signaling pathways in host cells and speculate how this might affect the outcome of Toxoplasma encephalitis.

DOI: https://doi.org/10.1038/s41420-023-01742-1
Int J Parasitol Drugs Drug Resist. 2023 Nov 24. pii: S2211-3207(23)00037-4. [Epub ahead of print]23 120-129

Evaluation of topotecan and 10-hydroxycamptothecin on Toxoplasma gondii: Implications on baseline DNA damage and repair efficiency.

Constanza Cristaldi, Ana M Saldarriaga Cartagena, Agustina Ganuza, William J Sullivan, Sergio O Angel, Laura Vanagas.

  Toxoplasma gondii is an obligate intracellular parasite in the phylum Apicomplexa that causes toxoplasmosis in humans and animals worldwide. Despite its prevalence, there is currently no effective vaccine or treatment for chronic infection. Although there are therapies against the acute stage, prolonged use is toxic and poorly tolerated. This study aims to explore the potential of repurposing topotecan and 10-hydroxycamptothecin (HCPT) as drugs producing double strand breaks (DSBs) in T. gondii. DSBs are mainly repaired by Homologous Recombination Repair (HRR) and Non-Homologous End Joining (NHEJ). Two T. gondii strains, RHΔHXGPRT and RHΔKU80, were used to compare the drug's effects on parasites. RHΔHXGPRT parasites may use both HRR and NHEJ pathways but RHΔKU80 lacks the KU80 protein needed for NHEJ, leaving only the HRR pathway. Here we demonstrate that topotecan and HCPT, both topoisomerase I venoms, affected parasite replication in a concentration-dependent manner. Moreover, variations in fluorescence intensity measurements for the H2A.X phosphorylation mark (γH2A.X), an indicator of DNA damage, were observed in intracellular parasites under drug treatment conditions. Interestingly, intracellular replicative parasites without drug treatment show a strong positive staining for γH2A.X, suggesting inherent DNA damage. Extracellular (non-replicating) parasites did not exhibit γH2A.X staining, indicating that the basal level of DNA damage is likely to be associated with replicative stress. A high rate of DNA replication stress possibly prompted the evolution of an efficient repair machinery in the parasite, making it an attractive target. Our findings show that topoisomerase 1 venoms are effective antiparasitics blocking T. gondii replication.

Keywords:  10-Hydroxycamptothecin; DNA damage; DNA damage response; Stress replication; Topotecan; Toxoplasma gondii

DOI:  https://doi.org/10.1016/j.ijpddr.2023.11.004
Microb Pathog. 2023 Dec 03. pii: S0882-4010(23)00517-X. [Epub ahead of print] 106484

Proteomics analysis of Toxoplasma gondii merozoites reveals regulatory proteins involved in sexual reproduction.

Guihua Zhao, Hongjie Dong, Lisha Dai, Huanhuan Xie, Hang Sun, Junmei Zhang, Qi Wang, Chao Xu, Kun Yin.

  Sexual reproduction plays a crucial role in the transmission and life cycle of toxoplasmosis. The merozoites are the only developmental stage capable of differentiation into male and female gametes, thereby initiating sexual reproduction to form oocysts that are excreted into the environment. Hence, our study aimed to perform proteomic analyses of T. gondii Pru strain merozoites, a pre-sexual developmental stage in cat IECs, and tachyzoites, an asexual developmental stage, using the tandem mass tag (TMT) method in order to identify the differentially expressed proteins (DEPs) of merozoites. Proteins functions were subjected to cluster analysis, and DEPs were validated through the qPCR method. The results showed that a total of 106 proteins were identified, out of which 85 proteins had quantitative data. Among these, 15 proteins were differentially expressed within merozoites, with four exhibiting up-regulation and being closely associated with the material and energy metabolism as well as the cell division of T. gondii. Two novel DEPs, namely S8GHL5 and A0A125YP41, were identified, and their homologous family members have been demonstrated to play regulatory roles in oocyte maturation and spermatogenesis in other species. Therefore, they may potentially exhibit regulatory functions during the differentiation of micro- and macro-gametophytes at the initiation stage of sexual reproduction in T. gondii. In conclusion, our results showed that the metabolic and divisional activities in the merozoites surpass those in the tachyzoites, thereby providing structural, material, and energetic support for gametophytes development. The discovery of two novel DEPs associated with sexual reproduction represents a significant advancement in understanding Toxoplasma sexual reproduction initiation and oocyst formation.

Keywords:  Definitive host; Merozoites; Sexual reproduction; TMT proteomics; Toxoplasma gondii

DOI:  https://doi.org/10.1016/j.micpath.2023.106484
Parasites Hosts Dis. 2023 Nov;61(4): 418-427

The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques.

Hae-Ji Kang, Jie Mao, Min-Ju Kim, Keon-Woong Yoon, Gi-Deok Eom, Ki-Back Chu, Eun-Kyung Moon, Fu-Shi Quan.

  Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

Keywords:  B1 gene; BALB/c mice; IgG; Toxoplasma gondii ME49

DOI:  https://doi.org/10.3347/PHD.23048
Geburtshilfe Frauenheilkd. 2023 Dec;83(12): 1431-1445

Toxoplasma gondii Infection in Pregnancy - Recommendations of the Working Group on Obstetrics and Prenatal Medicine (AGG - Section on Maternal Disorders).

AGG Section Maternal Diseases

  Aim The AGG (Working Group for Obstetrics and Prenatal Diagnostics, Section Maternal Diseases) has issued these recommendations to improve the detection and management of Toxoplasma gondii infection in pregnancy. Methods Members of the Task Force developed the recommendations and statements presented here using recently published literature. The recommendations were adopted after a consensus process by members of the working group. Recommendations This article focuses on the epidemiology and pathophysiology of Toxoplasma gondii infection in pregnancy and includes recommendations for maternal and fetal diagnosis, transmission prophylaxis, therapy, prevention, screening, and peripartum management.

Keywords:  Toxoplasma gondii; congenital toxoplasmosis; pregnancy; therapy; transmission

DOI:  https://doi.org/10.1055/a-2111-7394
BMC Biol. 2023 Dec 05. 21(1): 279

Plasmodium yoelii iron transporter PyDMT1 interacts with host ferritin and is required in full activity for malarial pathogenesis.

Mengjiao Zhong, Bing Zhou.

   BACKGROUND: The rapid reproduction of malaria parasites requires proper iron uptake. However, the process of iron absorption by parasites is rarely studied. Divalent metal transporter (DMT1) is a critical iron transporter responsible for uptaking iron. A homolog of human DMT1 exists in the malaria parasite genome, which in Plasmodium yoelii is hereafter named PyDMT1.
RESULTS: PyDMT1 knockout appears to be lethal. Surprisingly, despite dwelling in an iron-rich environment, the parasite cannot afford to lose even partial expression of PyDMT1; PyDMT1 hypomorphs were associated with severe growth defects and quick loss of pathogenicity. Iron supplementation could completely suppress the defect of the PyDMT1 hypomorph during in vitro culturing. Genetic manipulation through host ferritin (Fth1) knockout to increase intracellular iron levels enforced significant growth inhibition in vivo on the normal parasites but not the mutant. In vitro culturing with isolated ferritin knockout mouse erythrocytes completely rescued PyDMT1-hypomorph parasites.
CONCLUSION: A critical iron requirement of malaria parasites at the blood stage as mediated by this newly identified iron importer PyDMT1, and the iron homeostasis in malarial parasites is finely tuned. Tipping the iron balance between the parasite and host will efficiently kill the pathogenicity of the parasite. Lastly, PyDMT1 hypomorph parasites were less sensitive to the action of artemisinin.

Keywords:  Artemisinin; Ferritin; Iron; Plasmodium yoelli; PyDMT1

DOI:  https://doi.org/10.1186/s12915-023-01776-y
PLoS Pathog. 2023 Dec 04. 19(12): e1011818

Plasmodium falciparum utilizes pyrophosphate to fuel an essential proton pump in the ring stage and the transition to trophozoite stage.

Omobukola Solebo, Liqin Ling, Ikechukwu Nwankwo, Jing Zhou, Tian-Min Fu, Hangjun Ke.

During asexual growth and replication cycles inside red blood cells, the malaria parasite Plasmodium falciparum primarily relies on glycolysis for energy supply, as its single mitochondrion performs little or no oxidative phosphorylation. Post merozoite invasion of a host red blood cell, the ring stage lasts approximately 20 hours and was traditionally thought to be metabolically quiescent. However, recent studies have shown that the ring stage is active in several energy-costly processes, including gene transcription, protein translation, protein export, and movement inside the host cell. It has remained unclear whether a low glycolytic flux alone can meet the energy demand of the ring stage over a long period post invasion. Here, we demonstrate that the metabolic by-product pyrophosphate (PPi) is a critical energy source for the development of the ring stage and its transition to the trophozoite stage. During early phases of the asexual development, the parasite utilizes Plasmodium falciparum vacuolar pyrophosphatase 1 (PfVP1), an ancient pyrophosphate-driven proton pump, to export protons across the parasite plasma membrane. Conditional deletion of PfVP1 leads to a delayed ring stage that lasts nearly 48 hours and a complete blockage of the ring-to-trophozoite transition before the onset of parasite death. This developmental arrest can be partially rescued by an orthologous vacuolar pyrophosphatase from Arabidopsis thaliana, but not by the soluble pyrophosphatase from Saccharomyces cerevisiae, which lacks proton pumping activities. Since proton-pumping pyrophosphatases have been evolutionarily lost in human hosts, the essentiality of PfVP1 suggests its potential as an antimalarial drug target. A drug target of the ring stage is highly desired, as current antimalarials have limited efficacy against this stage.

DOI: https://doi.org/10.1371/journal.ppat.1011818
Food Microbiol. 2024 Apr;pii: S0740-0020(23)00190-9. [Epub ahead of print]118 104403

Whole genome enrichment approach for genomic surveillance of Toxoplasma gondii.

Balaji Sundararaman, Karen Shapiro, Andrea Packham, Lauren E Camp, Rachel S Meyer, Beth Shapiro, Richard E Green.

  Pathogenic bacteria, viruses, fungi, and protozoa can cause food and waterborne diseases. Surveillance methods must therefore screen for these pathogens at various stages of water distribution and of food from production to consumption. Detection using nucleic acid amplification methods offer rapid identification, but such methods have limited utility for characterizing populations, variant types or virulence traits of pathogens. Whole genome sequencing (WGS) can be used to determine this information. However, pathogens must be isolated and cultured to yield sufficient DNA for WGS, which is laborious or not feasible for certain stages of parasites like oocysts of Toxoplasma gondii. We previously developed the Circular Nucleic acid Enrichment Reagent (CNER) method to make whole genome enrichment (WGE) baits for difficult-to-grow bacterial pathogens. WGE using CNERs facilitates direct sequencing of pathogens from samples without the need to isolate and grow them. Here, we made WGE-CNERs for T. gondii to demonstrate the use of the CNER method to make baits to enrich the large genomes of water and foodborne protozoan pathogens. By sequencing, we detected as few as 50 parasites spiked in an oyster hemolymph matrix. We discuss the use of WGE-CNERs for genomic surveillance of food and waterborne pathogens.

Keywords:  Genomic epidemiology; One health; Pathogen surveillance; Shellfish; Toxoplasmosis; Whole genome enrichment

DOI:  https://doi.org/10.1016/j.fm.2023.104403
Int J Parasitol Drugs Drug Resist. 2023 Nov 24. pii: S2211-3207(23)00040-4. [Epub ahead of print]23 106-119

Effects of the G-quadruplex-binding drugs quarfloxin and CX-5461 on the malaria parasite Plasmodium falciparum.

Holly M Craven, Guilherme Nettesheim, Pietro Cicuta, Andrew M Blagborough, Catherine J Merrick.

  Plasmodium falciparum is the deadliest causative agent of human malaria. This parasite has historically developed resistance to most drugs, including the current frontline treatments, so new therapeutic targets are needed. Our previous work on guanine quadruplexes (G4s) in the parasite's DNA and RNA has highlighted their influence on parasite biology, and revealed G4 stabilising compounds as promising candidates for repositioning. In particular, quarfloxin, a former anticancer agent, kills blood-stage parasites at all developmental stages, with fast rates of kill and nanomolar potency. Here we explored the molecular mechanism of quarfloxin and its related derivative CX-5461. In vitro, both compounds bound to P. falciparum-encoded G4 sequences. In cellulo, quarfloxin was more potent than CX-5461, and could prevent establishment of blood-stage malaria in vivo in a murine model. CX-5461 showed clear DNA damaging activity, as reported in human cells, while quarfloxin caused weaker signatures of DNA damage. Both compounds caused transcriptional dysregulation in the parasite, but the affected genes were largely different, again suggesting different modes of action. Therefore, CX-5461 may act primarily as a DNA damaging agent in both Plasmodium parasites and mammalian cells, whereas the complete antimalarial mode of action of quarfloxin may be parasite-specific and remains somewhat elusive.

Keywords:  CX-5461; G-quadruplex; Malaria; Plasmodium; quarfloxin; repositioning

DOI:  https://doi.org/10.1016/j.ijpddr.2023.11.007
Front Cell Infect Microbiol. 2023 ;13 1334002

Editorial: Lipids in host and protozoan parasite interaction.

Silvia Haase, Lesly Temesvari, Kai Zhang.



Keywords:  cell signaling; lipid droplets; lipid metabolism; lipid rafts; membrane trafficking; parasites

DOI:  https://doi.org/10.3389/fcimb.2023.1334002
Epigenomics. 2023 Dec 07.

Histone deacetylase 6: at the interface of cancer and neurodegeneration.

Jingjing Pu, Amit Sharma, Jian Hou, Ingo Gh Schmidt-Wolf.

  With the recognition in the early 1960s that histones can be post-translationally modified, the list of different post-translational modifications of histones and their biological consequences has continued to expand. In addition, the idea of the 'histone code' hypothesis, later introduced by David Allis and colleagues, further broaden the horizon of chromatin biology. Currently, there is a wealth of knowledge about the transition between the active and the repressive state of chromatin, and modifications of histones remains at the center of chromatin biology. Histone deacetylases (HDACs) in particular are of great importance for the therapeutic success of cancer treatment. Focusing primarily on HDAC6, herein we have briefly highlighted its unique involvement in cancer and also apparently in neurodegeneration.

Keywords:  cancer; epigenetics; histone deacetylase; neurodegeneration; therapy

DOI:  https://doi.org/10.2217/epi-2023-0373
Cell Metab. 2023 Dec 05. pii: S1550-4131(23)00417-5. [Epub ahead of print]35(12): 2097-2099

mTOR takes charge: Relaying uncharged tRNA levels by mTOR ubiquitination.

Estela Jacinto.

Nutrient availability is conveyed to the mechanistic target of rapamycin (mTOR), which couples metabolic processes with cell growth and proliferation. How mTOR itself is modulated by amino acid levels remains poorly understood. Ge and colleagues now demonstrate that broad sensing of uncharged tRNAs by GCN2/FBXO22 inactivates mTOR complex 1 (mTORC1) via mTOR ubiquitination.

DOI: https://doi.org/10.1016/j.cmet.2023.11.006
Biomed Pharmacother. 2023 Dec 02. pii: S0753-3322(23)01749-3. [Epub ahead of print]170 115951

Drug addiction and treatment: An epigenetic perspective.

Arunkumar Singh Koijam, Kabrambam Dasanta Singh, Bunindro Singh Nameirakpam, Reena Haobam, Yallappa Rajashekar.

  Drug addiction is a complex disease affected by numerous genetic and environmental factors. Brain regions in reward pathway, neuronal adaptations, genetic and epigenetic interactions causing transcriptional enhancement or repression of multiple genes induce different addiction phenotypes for varying duration. Addictive drug use causes epigenetic alterations and similarly epigenetic changes induced by environment can promote addiction. Epigenetic mechanisms include DNA methylation and post-translational modifications like methylation, acetylation, phosphorylation, ubiquitylation, sumoylation, dopaminylation and crotonylation of histones, and ADP-ribosylation. Non-coding RNAs also induce epigenetic changes. This review discusses these above areas and stresses the need for exploring epidrugs as a treatment alternative and adjunct, considering the limited success of current addiction treatment strategies. Epigenome editing complexes have lately been effective in eukaryotic systems. Targeted DNA cleavage techniques such as CRISPR-Cas9 system, CRISPR-dCas9 complexes, transcription activator-like effector nucleases (TALENs) and zinc-finger nucleases (ZFNs) have been exploited as targeted DNA recognition or anchoring platforms, fused with epigenetic writer or eraser proteins and delivered by transfection or transduction methods. Efficacy of epidrugs is seen in various neuropsychiatric conditions and initial results in addiction treatment involving model organisms are remarkable. Epidrugs present a promising alternative treatment for addiction.

Keywords:  Addiction; CRISPR-Cas9; Epigenetic; Epigenome editing complexes; TALEN; ZFN

DOI:  https://doi.org/10.1016/j.biopha.2023.115951
Nucleic Acids Res. 2023 Dec 07. pii: gkad1172. [Epub ahead of print]

Lysine acetylation regulates the AT-rich DNA possession ability of H-NS.

Yabo Liu, Mengqing Zhou, Yifan Bu, Liang Qin, Yuanxing Zhang, Shuai Shao, Qiyao Wang.

H-NS, the histone-like nucleoid-structuring protein in bacteria, regulates the stability of the bacterial genome by inhibiting the transcription of horizontally transferred genes, such as the type III and type VI secretion systems (T3/T6SS). While eukaryotic histone posttranslational modifications (PTMs) have been extensively studied, little is known about prokaryotic H-NS PTMs. Here, we report that the acetylation of H-NS attenuates its ability to silence horizontally transferred genes in response to amino acid nutrition and immune metabolites. Moreover, LC-MS/MS profiling showed that the acetyllysine sites of H-NS and K120 are indispensable for its DNA-binding ability. Acetylation of K120 leads to a low binding affinity for DNA and enhances T3/T6SS expression. Furthermore, acetylation of K120 impairs the AT-rich DNA recognition ability of H-NS. In addition, lysine acetylation in H-NS modulates in vivo bacterial virulence. These findings reveal the mechanism underlying H-NS PTMs and propose a novel mechanism by which bacteria counteract the xenogeneic silencing of H-NS.

DOI: https://doi.org/10.1093/nar/gkad1172
bioRxiv. 2023 Nov 26. pii: 2023.11.26.568742. [Epub ahead of print]

Learning sequence, structure, and function representations of proteins with language models.

Tymor Hamamsy, Meet Barot, James T Morton, Martin Steinegger, Richard Bonneau, Kyunghyun Cho.

The sequence-structure-function relationships that ultimately generate the diversity of extant observed proteins is complex, as proteins bridge the gap between multiple informational and physical scales involved in nearly all cellular processes. One limitation of existing protein annotation databases such as UniProt is that less than 1% of proteins have experimentally verified functions, and computational methods are needed to fill in the missing information. Here, we demonstrate that a multi-aspect framework based on protein language models can learn sequence-structure-function representations of amino acid sequences, and can provide the foundation for sensitive sequence-structure-function aware protein sequence search and annotation. Based on this model, we introduce a multi-aspect information retrieval system for proteins, Protein-Vec, covering sequence, structure, and function aspects, that enables computational protein annotation and function prediction at tree-of-life scales.

DOI: https://doi.org/10.1101/2023.11.26.568742