Dataset Information

Arginine and lysine methylproteomes of Giardia duodenalis

ABSTRACT: Methylation is a common post-translational modification of lysine and arginine in eukaryotic proteins. To date, these methylomes are best characterised in model organisms and higher eukaryotes. Herein, we integrated bioinformatics, proteomics and high-content drug-screening to comprehensively explore protein methylation in the human gastrointestinal parasite and deep-branching eukaryote, Giardia duodenalis. We demonstrate Giardia and other Diplomonadida species lack arginine-methyltransferases and have remodelled RGG/RG motifs preferred by these enzymes. Further, Giardia had no detectable methylarginine in vitro, demonstrating the first eukaryote with no arginine methylome. In contrast, we performed detailed curation of 11 putative lysine-methyltransferases, including highly-diverged SET-domain proteins, and novel annotations demonstrating conserved eEF1a methyllysine. We identified >200 high-confidence methyllysine sites, highlighting methyllysine within coiled-coil features, and for Giardia cytoskeletal regulation. Lastly, using known methylation-inhibitors, we demonstrate inhibition of methylation plays a key role in replication and cyst formation in this parasite.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Giardia Lamblia Atcc 50803

TISSUE(S): Trophozoite, Binucleate Cell

DISEASE(S): Giardiasis

SUBMITTER: Samantha Emery

LAB HEAD: Aaron R Jex

PROVIDER: PXD016813 | Pride | 2020-08-07

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	GiardiaKMe-DimethylKSites.txt	Txt
	GiardiaKMe-MethylKSites.txt	Txt
	GiardiaKMe-TrimethylKSites.txt	Txt
	GiardiaKMe-msms.txt	Txt
	GiardiaKMe-peptides.txt	Txt

Items per page:

1 - 5 of 45

Publications

Eukaryote-Conserved Methylarginine Is Absent in Diplomonads and Functionally Compensated in Giardia.

Emery-Corbin Samantha J SJ Hamey Joshua J JJ Ansell Brendan R E BRE Balan Balu B Tichkule Swapnil S Stroehlein Andreas J AJ Cooper Crystal C McInerney Bernie V BV Hediyeh-Zadeh Soroor S Vuong Daniel D Crombie Andrew A Lacey Ernest E Davis Melissa J MJ Wilkins Marc R MR Bahlo Melanie M Svärd Staffan G SG Gasser Robin B RB Jex Aaron R AR

Molecular biology and evolution 20201201 12

Methylation is a common posttranslational modification of arginine and lysine in eukaryotic proteins. Methylproteomes are best characterized for higher eukaryotes, where they are functionally expanded and evolved complex regulation. However, this is not the case for protist species evolved from the earliest eukaryotic lineages. Here, we integrated bioinformatic, proteomic, and drug-screening data sets to comprehensively explore the methylproteome of Giardia duodenalis-a deeply branching parasiti ...[more]

PMID: 32702104

Similar Datasets

Project description:Giardia causes more episodes of illness worldwide than any other parasite. It is a flagellated cyst-forming enteric pathogen that inhabits the lumen of the small intestine and although it is clear that more and less virulent strains do occur, pathogenesis and virulence of Giardia remains poorly understood. Identification of Giardia virulence factors is highly desirable since their expression would be the best indicator for determining disease outcome. The advent of quantitative proteomics has been timely in investigating which proteins are likely to be secreted virulence factors. Here we successfully apply such analyses to the whole parasite and to the supernatants derived from the parasite, in order to ascertain a repertoire of secreted proteins. In our initial analysis we compared replicates of Giardia cells and culture supernatants from the two major lineages which cause human disease – assemblage A and assemblage B. The genome of Giardia is believed to contain open reading frames which could encode as many as 6000 proteins. In our study we confirm expression of ~1600 proteins from each assemblage, the vast majority of which being common to both lineages. To look for actual enrichment of secreted proteins, we considered the ratio of proteins in the supernatant compared with the pellet. This simple method defines a small group of enriched proteins, putatively secreted at a steady state by cultured growing Giardia trophozoites of both assemblages. This secretome contains a high proportion annotated to have N’ terminus signal peptides, with some showing evidence of having recently evolved under strong positive selective pressure. The most abundant secreted proteins include known virulence factors such as the Cathepsin B cysteine proteases and members of a Giardia superfamily of Cysteine Rich Proteins which comprises VSPs, HCMPs and a new class of virulence factors, the Giardia Tenascins. Since we also find that physiological function of human enteric epithelial cells can be disrupted by soluble factors even in the absence of the trophozoites we are able to propose a straightforward model of Giardia pathogenesis incorporating key roles for the major Giardia derived soluble mediators.