Dataset Information

BioID-based proteomic analysis of CARP3-proximal proteins in Trypanosoma brucei bloodstream forms

ABSTRACT: How flagellar signaling regulates the host interaction of parasites remains a challenge due to poor conservation of signaling systems with those in cilia of higher organisms. The trypanosome-specific cAMP response protein 3 (CARP3) shows developmentally regulated localization at the flagellar tip membrane, where it is essential for parasite swarming and colonization of the tsetse fly insect vector. This project describes a proximity-dependent proteomics approach (BioID) that identified proteins in close vicinity of CARP3 in the bloodstream stage of Trypanosoma brucei.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Trypanosoma Brucei

SUBMITTER: Dupuy Jean-William

LAB HEAD: Michael Boshart

PROVIDER: PXD025357 | Pride | 2022-08-17

REPOSITORIES: Pride

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Publications

A multi-adenylate cyclase regulator at the flagellar tip controls African trypanosome transmission.

Bachmaier Sabine S Giacomelli Giacomo G Calvo-Alvarez Estefanía E Vieira Larissa Rezende LR Van Den Abbeele Jan J Aristodemou Aris A Lorentzen Esben E Gould Matt K MK Brennand Ana A Dupuy Jean-William JW Forné Ignasi I Imhof Axel A Bramkamp Marc M Salmon Didier D Rotureau Brice B Boshart Michael M

Nature communications 20220916 1

Signaling from ciliary microdomains controls developmental processes in metazoans. Trypanosome transmission requires development and migration in the tsetse vector alimentary tract. Flagellar cAMP signaling has been linked to parasite social motility (SoMo) in vitro, yet uncovering control of directed migration in fly organs is challenging. Here we show that the composition of an adenylate cyclase (AC) complex in the flagellar tip microdomain is essential for tsetse salivary gland (SG) colonizat ...[more]

PMID: 36114198

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Project description:The bloodstream forms of Trypanosoma brucei (BSF), responsible for the sleeping sickness, primarily evaluate in the blood of its mammalian hosts. The skin and the adipose tissues were recently identified as additional major sites for the parasite proliferation and transmission through its insect vector blood ingestion. Glucose is the only carbon source used by the blood parasites to feed its central metabolism, however, the metabolic behavior of the tissue-adapted parasites has not yet been addressed. Since production of glycerol is an important primary function of adipocytes, we have adapted the BSF trypanosomes to glucose-depleted and glycerol-rich growth medium (CMM_Gly/GlcNAc) and compared its metabolism and proteome with standard glucose-rich growth conditions (CMM_Glu). BSF consume 2-times more oxygen per carbon consumed in CMM_Gly/GlcNAc and is 11.5-times more sensitive to SHAM, a specific inhibitor of the alternative oxidase, which is consistent with the expected absolute requirement of the mitochondrial respiratory chain activity to convert glycerol into dihydroxyacetone phosphate. Metabolomic analyses by mass spectrometry showed that 13C-labeled glycerol is incorporated into hexose phosphates through gluconeogenesis. After several weeks of growth in CMM_Gly/GlcNAc, BSF adapt their metabolism by increasing and decreasing the rate of glycerol and glucose consumption, respectively. As expected RNAi-mediated down-regulation of glycerol kinase expression abolished glycerol degradation and is lethal for BSF growth in CMM_Gly/GlcNAc. However, the glycerol kinase activity is 7.8-fold decreased in CMM_Gly/GlcNAc, as confirms by western blotting and proteomic analyses, which suggests that the huge excess of glycerol kinase, which is not necessary for glycerol metabolism, is required for another yet undetermined non-essential function in glucose rich-conditions. Altogether these data shows that BSF trypanosomes are well adapted to glycerol-rich conditions, which could be encountered by the parasite in extravascular niches, such as skin and adipose tissues.

			Action	DRS
	Link_between_files.xlsx	Xlsx
	TriTrypDB-51_TbruceiTREU927_AnnotatedProteins.fasta	Fasta
	andromeda.zip	Other
	checksum.txt	Txt
	mb161027_01.raw	Raw