PXD014998

PXD014998 is an original dataset announced via ProteomeXchange.

Title	Protein ubiquitylation is essential for the schizont to merozoite transition in Plasmodium falciparum blood-stage development
Description	Ubiquitylation is a common post translational modification of eukaryotic proteins. Protein ubiquitylation increases in the transition from intracellular schizont to extracellular merozoite in the asexual blood stage cycle of the human malaria parasite, Plasmodium falciparum (Pf). Here, we identify specific ubiquitylation sites of protein substrates in three intracellular parasite stages and extracellular merozoites; a total of 1464 sites in 546 proteins were identified. 469 ubiquitylated proteins were identified in merozoites compared with only 160 in the preceding intracellular schizont stage, indicating a large increase in protein ubiquitylation associated with merozoite maturation. Following merozoite invasion of erythrocytes, few ubiquitylated proteins were detected in the first intracellular ring stage but as parasites matured through trophozoite to schizont stages the extent of ubiquitylation increased. We identified commonly used ubiquitylation motifs and groups of ubiquitylated proteins in specific areas of cellular function, for example merozoite pellicle proteins involved in erythrocyte invasion, exported proteins, and histones. To investigate the importance of ubiquitylation we screened ubiquitin pathway inhibitors in a parasite growth assay and identified the ubiquitin activating enzyme (UBA1 or E1) inhibitor, MLN7243 (TAK-243) to be particularly effective. This small molecule was shown to be a potent inhibitor of recombinant PfUBA1, and a structural homology model of MLN7243 bound to the parasite enzyme highlights avenues for the development of P. falciparum specific inhibitors. We constructed a genetically modified parasite with a rapamycin-inducible functional deletion of uba1; addition of either MLN7243 or rapamycin to the recombinant parasite line resulted in the same phenotype, with parasite development blocked at the schizont stage. These results indicate that the intracellular target of MLN7243 is UBA1, and this activity is essential for the final differentiation of schizonts to merozoites. The ubiquitylation of many merozoite proteins and their disappearance in ring stages are consistent with the idea that ubiquitylation leads to their destruction via the proteasome once their function is complete following invasion, which would allow amino acid recycling in the period prior to the parasite’s elaboration of a new food vacuole.
HostingRepository	PRIDE
AnnounceDate	2020-05-20
AnnouncementXML	Submission_2020-08-11_05:49:07.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Vikram Sharma
SpeciesList	scientific name: Plasmodium falciparum; NCBI TaxID: 5833;
ModificationList	ubiquitination signature dipeptidyl lysine
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2019-08-12 07:30:44	ID requested
1	2020-05-20 02:08:52	announced
⏵ 2	2020-08-11 05:49:08	announced	2020-08-11: Updated publication reference for PubMed record(s): 32569299.

Green JL, Wu Y, Encheva V, Lasonder E, Prommaban A, Kunzelmann S, Christodoulou E, Grainger M, Truongvan N, Bothe S, Sharma V, Song W, Pinzuti I, Uthaipibull C, Srichairatanakool S, Birault V, Langsley G, Schindelin H, Stieglitz B, Snijders AP, Holder AA, Ubiquitin activation is essential for schizont maturation in Plasmodium falciparum blood-stage development. PLoS Pathog, 16(6):e1008640(2020) [pubmed]

submitter keyword: Proteomics, Plasmodium, Malaria, Ubiquitylation

Anthony A. Holder
contact affiliation	Senior Group Leader, Malaria Parasitology Laboratory The Francis Crick Institute 1 Midland Road London NW1 1AT
contact email	Tony.Holder@crick.ac.uk
lab head
Vikram Sharma
contact affiliation	Plymouth University Peninsula Schools of Medicine and Dentistry
contact email	vikram.sharma@plymouth.ac.uk
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/05/PXD014998

PRIDE project URI

• PRIDE
  1. PXD014998
     1. Label: PRIDE project
     2. Name: Protein ubiquitylation is essential for the schizont to merozoite transition in Plasmodium falciparum blood-stage development