Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Tryptophan C-mannosylation is critical for Plasmodium falciparum transmission Dataset1

ABSTRACT: C-mannosylation stabilizes proteins bearing a thrombospondin repeat (TSR) domain in metazoans. Here we show that Plasmodium falciparum expresses a DPY19 C-mannosyltransferase in the endoplasmic reticulum and that DPY19-deficiency abolishes C-glycosylation, destabilizes members of the TRAP adhesin family and inhibits transmission to mosquitoes. P. falciparum gametogenesis was imaged in its entirety in four dimensions using lattice light-sheet microscopy. This revealed defects in egress and exflagellation for DPY19 microgametes. While exflagellation was diminished, DPY19 microgametes still fertilized macrogametes, forming ookinetes but these were abrogated for mosquito infection. The gametogenesis defects corresponded with destabilization of MTRAP, which we show is C-mannosylated in P. falciparum, and the ookinete defect was concordant with defective CTRP secretion on the DPY19 background. Genetic complementation of DPY19 restored ookinete infectivity, sporozoite production and C-mannosylation activity. Therefore, tryptophan C-mannosylation by DPY19 in the early secretory pathway ensures TSR protein quality control at two lifecycle stages for successful transmission of the human malaria parasite.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Plasmodium Falciparum

TISSUE(S): Blood

SUBMITTER: Nichollas Scott

LAB HEAD: Justin A. Boddey

PROVIDER: PXD033470 | Pride | 2022-07-06

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	20220211_NSco-3748_EGB_mtrap_CMT.raw	Raw
	20220211_NSco-3748_EGB_mtrap_CMT_PRM.raw	Raw
	20220211_NSco-3748_EGB_mtrap_WT.raw	Raw
	20220211_NSco-3748_EGB_spatr_CMT.raw	Raw
	20220211_NSco-3748_EGB_spatr_CMT_PRM.raw	Raw

Items per page:

1 - 5 of 10

Similar Datasets

Discovery and characterization of a new class of O-linking oligosaccharyltransferase from bacteria in the Moraxellaceae family

Project description:Bacterial protein glycosylation can be mediated by oligosaccharyltransferases (OTases) that transfer a preassembled lipid-linked oligosaccharide or polysaccharide en bloc to acceptor proteins. O-linking OTases transfer O-antigen or capsular polysaccharides to the side chains of serine or threonine residues. Three major families of bacterial O-linking OTases have been described so far: PglL, PglS, and TfpO. TfpO enzymes are limited to transferring only short glycans whereas there are no clear upper limits for the other two families. Herein, we describe the discovery of a novel family of bacterial O-linking OTases from Moraxellacea bacteria that are similar in size and sequence to TfpO enzymes but can transfer long-chain bacterial glycans to acceptor proteins. Bioinformatic analyses show that these enzymes cluster in different clades than known bacterial OTases. Using a representative enzyme from Moraxella osloensis termed TfpMMo, we determine that the enzyme glycosylates the C-terminal amino acid side chain of a pilin protein and find that pilin fragments as short as three amino acids are substrates for the OTase. The ability of TfpMMo to transfer long-chain polysaccharide shows that this ability is not limited to the PglS and PglL families. TfpMMo is also shown to have broad substrate specificity and can transfer diverse glycans including those with glucose, galactose, or 2-N-acetyl sugars at the reducing end. The glycan substrate promiscuity of TfpMMo could allow this enzyme to be used to produce bacterial glycoconjugate vaccines. The discovery of a new class of O-linking OTase furthers our understanding of the mechanisms that underly glycan specificity by these and other O-linking OTases and enables more comparative studies of this important enzyme family.

2022-10-13 | PXD033468 | Pride

Anopheles gambiae immune responses to human and rodent Plasmodium parasite species

Project description:Transmission of malaria is dependent on the successful completion of the Plasmodium lifecycle in the Anopheles vector. Major obstacles are encountered in the midgut tissue, where most parasites are killed by the mosquito’s immune system. In the present study, DNA microarray analyses have been used to compare Anopheles gambiae responses to invasion of the midgut epithelium by the ookinete stage of the human pathogen Plasmodium falciparum and the rodent experimental model pathogen P. berghei. Invasion by P. berghei had a more profound impact on the mosquito transcriptome, including a variety of functional gene classes, while P. falciparum elicited a broader immune response at the gene transcript level. Ingestion of human malaria-infected blood lacking invasive ookinetes also induced a variety of immune genes, including several anti-Plasmodium factors. By comparing gene expression patterns between carcassess or guts of mosquitoes that fed on a P. falciparum or P. berghei wt and mosquitoes that fed on invasion incapable strains we gain information on the A. gambiae transcriptional responses to the invading ookinete at 24 hours after feeding. By comparing gene expression patterns between carcassess or guts of mosquitoes that fed on a P. falciparum ookinete invasion incapable strain and mosquitoes that fed on non-infected blood we gain information on the A. gambiae transcriptional responses to malaria infected blood in absence of ookinete invasion at 24 hours after feeding. By comparing gene expression patterns between mosquitoes at 4 hours after being injected with either E. coli or S. aureus and mosquitoes injected with sterile PBS we gain information on the mosquito's transcriptional response to these bacterial challenges.

2007-08-15 | E-GEOD-8789 | biostudies-arrayexpress

Anopheles gambiae immune responses to human and rodent Plasmodium parasite species

2007-08-15 | GSE8789 | GEO

Phosphoproteomic analysis of activated P. falciparum gametocytes, wild type NF54 and PfCDPK4 knockout

Project description:Gametocytes of the malaria parasite Plasmodium are taken up by the mosquito vector with an infectious blood meal, representing a critical stage for parasite transmission. It is well known that calcium dependent protein kinases (CDPKs) play key roles in calcium-mediated signaling across the complex life cycle of the parasite and we sought to understand their role in transmission from the mammalian host to the mosquito vector. Here, we investigated the role of CDPK4 in the life cycle of the human-infective parasite Plasmodium falciparum . We created Plasmodium falciparum cdpk4 knockout parasites by targeted gene deletion, and this deletion had no effect on blood stage development or gametocyte development. However, cdpk4 knockout parasites showed a severe defect in male gametogenesis and the emergence of male flagellated gametes. To understand this defect, we performed mass spectrometry-based phosphoproteomic analysis of wild type and Plasmodium falciparum cdpk4 knockout late gametocyte stages in order to identify key CDPK4-mediated phosphorylation events that may be important for the regulation of male gametogenesis.

2021-06-03 | MSV000087569 | MassIVE

A seven-helix protein acts as a key regulator of translation during human-to-mosquito transmission of Plasmodium falciparum

Project description:Plasmodium falciparum is a unicellular parasite responsible for the majority of 440,000 death due to malaria every year. Due to their essential role for malaria transmission, gametocytes represent prime targets for transmission-blocking strategies intended to prevent spread of the deadly disease. In this study, we explored the signaling pathways leading to gametogenesis and identified a hitherto unknown protein, which structurally belongs to the class of seven-helix proteins and which thus was termed 7-helix-1. The protein is specifically expressed in female gametocytes and gene disruption leads to impaired gamete formation and thus reduced transmission of malaria parasites to mosquitoes. The loss of 7-helix-1 caused significant changes in the expression of components of the molecular machinery needed by eukaryotic cells to synthesize proteins. We thus propose that 7-helix-1 is a key regulator needed to coordinate the increased need of proteins at the onset of gametogenesis.

2018-02-23 | GSE103012 | GEO

PRM analysis of C-glycosylation substrate

Project description:To assess the impact of mutations on CMT activity C-glycosylation within RNase E was monitored using PRM analysis.

2022-11-01 | PXD032391 | Pride

Single-cell transcriptomics to define Plasmodium falciparum stage-transition in the mosquito midgut

Project description:Malaria inflicts the highest rate of morbidity and mortality among the vector-borne diseases. The dramatic bottleneck of parasite numbers that occurs in the gut of the obligatory mosquito vector provides a promising target for novel control strategies. Using single-cell transcriptomics, we analyzed Plasmodium falciparum development in the mosquito gut, from unfertilized female gametes through the first 20 hours post blood feeding, including the zygote and ookinete stages. This study revealed the temporal gene expression of the ApiAP2 family of transcription factors, and of parasite stress genes in response to the harsh environment of the mosquito midgut. Further, employing structural protein prediction analyses we found several upregulated genes predicted to encode intrinsically disordered proteins (IDPs), a category of proteins known for their importance in regulation of transcription, translation and protein-protein interactions. IDPs are known for their antigenic properties and may serve as suitable targets for antibody or peptide-based transmission suppression strategies. In total, this study uncovers the P. falciparum transcriptome from early-to-late parasite development in the mosquito midgut, inside its natural vector, which provides an important resource for future malaria transmission-blocking initiatives. Single-cell data can be visualized interactively via https://mubasher-mohammed.shinyapps.io/shinyapp/ In-house bash, R code scripts and data that were implemented in this study are available on GitHub https://github.com/ANKARKLEVLAB/Single-cell-P.falciparum-midgut .

2023-02-16 | GSE222586 | GEO

Genome-wide RIP-Chip analysis of translational repressor-bound mRNAs in the Plasmodium gametocyte

Project description:Following fertilisation, translation of stored, but previously translationally repressed transcripts, is instrumental in shaping the proteome of the early embryo of multicellular eukaryotes; further development relies on de novo transcription of the zygotic genome. A related process was evidenced in the unicellular protozoa Plasmodium berghei during transmission to the mosquito host where fertilisation and zygote development result in the formation of an entirely new cell type, the ookinete. DOZI and CITH are mRNP components that maintain mRNAs silent in the female precursor cell. In the absence of DOZI or CITH, hundreds of transcripts are destabilised, which does not block fertilization upon transmission to the mosquito host, but impairs further development of the malaria parasite. However, how many transcripts are bound by DOZI/CITH in these mRNPs and what are they remain unknown. Here, we investigated the extent of maternal contribution during post-fertilization development of P. berghei by RNA immunoprecipitation (RIP)-Chip data mining and provided further functional significance using gene tagging and gene knockout approaches. We provide conclusive evidence that in the rodent malaria parasite Plasmodium berghei 50% of the transcriptome of the female sexual precursor cell is stably associated with the translational repressors DOZI and CITH allowing morphogenesis of the zygote into the ookinete to proceed in the absence of RNA polymerase II transcription. Using in situ tagging with GFP we find for the first time novel genes that rely on translational repression on the 5' UTR. This maternal repressome provides the developing ookinete with coding potential for key molecules required during developmental progression: a near complete set of factors required for the gliding motility apparatus, but also protein trafficking, adhesins, proteases and other factors.

2014-10-23 | E-MTAB-2900 | biostudies-arrayexpress

A multipass membrane protein interacts with the cGMP-dependent protein kinase to regulate critical calcium signals in malaria parasites

Project description:In malaria parasites, all cGMP-dependent signalling is mediated through a single cGMP-dependent protein kinase (PKG). A major function of PKG is to control calcium signals essential for the parasite to exit red blood cells or for transmission to the mosquito vector. However, how PKG controls these signals in the absence of known second messenger-dependent calcium channels or scaffolding proteins remains a mystery. Here we identify a tightly-associated PKG partner protein in both Plasmodium falciparum asexual blood stages and P. berghei gametocytes. Named ICM1, the partner is a polytopic membrane protein with homology to transporters and calcium channels, raising the possibility of a direct functional link between PKG and calcium homeostasis. Phosphoproteomic analyses in both Plasmodium species highlight a densely phosphorylated region of ICM1 with multiple phosphorylation events dependent upon PKG activity. Stage-specific depletion of ICM1 in P. berghei blocks gametogenesis due to the inability of mutant parasites to mobilise intracellular calcium upon PKG activation, whilst conditional disruption of P. falciparum ICM1 results in reduced cGMP-dependent calcium mobilisation associated with defective egress and invasion. Our findings provide new insights into the atypical calcium homeostasis in malaria parasites essential for pathology and disease transmission.

2021-02-10 | PXD020381 | Pride

Exploring the glycosylation state of the Cryptosporidium parvum CpTSP protein family-HILIC glycopeptide enrichment

Project description:Cryptosporidium parvum is a zoonotic apicomplexan parasite and a common cause of diarrheal disease worldwide. The development of vaccines to prevent or limit infection remains an important goal for tackling these diarrheal diseases, which are a significant cause of infant morbidity in the developing world. The only approved vaccine against an apicomplexan parasite targets conserved adhesins possessing a thrombospondin repeat (TSR) domains. Orthologous TSR domain-containing proteins are commonplace in the apicomplexa and C. parvum possess 12 such proteins. Here, we explore the molecular evolution and conservation of these proteins and examine their abundance in C. parvum oocysts to assess the likelihood that they may be useful as vaccine candidates. We go onto examine the glycosylation states of these proteins using antibody-enabled and ZIC-HILIC enrichment techniques, which revealed that these proteins are modified with C-linked Hex and N-linked Hex5-6HexNAc2 glycans.

2023-01-12 | PXD023936 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data