Project

Back to overview

Transporters of Trypanosoma brucei: Phylogeny - Physiology - Pharmacology

Applicant Mäser Pascal
Number 141913
Funding scheme Sinergia
Research institution Swiss Tropical and Public Health Institute Medical Services and Diagnostic Universität Basel
Institution of higher education University of Basel - BS
Main discipline Molecular Biology
Start/End 01.06.2013 - 28.02.2017
Approved amount 1'600'000.00
Show all

All Disciplines (4)

Discipline
Molecular Biology
Biochemistry
Cellular Biology, Cytology
Pharmacology, Pharmacy

Keywords (8)

ABC transporter; Saccharomyces cerevisiae; Trypanosoma brucei; Drug target; Lipid precursor; Xenopus laevis oocyte; Amino acid permease; Ion channel

Lay Summary (German)

Lead
Der Transport von Nährstoffen ist wichtig für jede Zelle, besonders aber für Parasiten wie Afrikanische Trypanosomen, welche sich im Blut ihrer Wirte vermehren und perfekt an diesen Lebensraum angepasst sind. Trypanosomen sind abhängig vom Import von Nährstoffen aus dem Blut. Wir untersuchen die dazu benötigten Transporter und testen, welche für die Parasiten essentiell sind und von pharmakologischem Interesse, sei es als Angriffspunkt für Wirkstoffe oder als Eintrittspforte.
Lay summary
Alle Zellen besitzen Transporter, durch die sie Nährstoffe aufnehmen und metabolische Endprodukte abgeben. Besonders wichtig sind solche Nährstofftransporter für Parasiten, denn die Aufnahme von Nährstoffen vom Wirtsorganismus gehört zum Wesen der parasitären Lebensweise. Neben ihren physiologischen Funktionen sind Transporter auch von grossem pharmakologischen Interesse, als Eintrittspforten für Wirkstoffe oder direkt als deren Angriffspunkte. In unserem Sinergia Projekt untersuchen wir den Erreger der Afrikanischen Schlafkrankheit, Trypanosoma brucei, und testen, welche Nährstofftransporter essentiell für den einzelligen Parasiten sind und welche Transporter auch Medikamente und andere Wirkstoffe transportieren können. Insbesondere untersuchen wir Ionenkanäle, Aminosäurenpermeasen, ABC Transporter (ABC steht für ATP-binding cassette) und Transporter von Lipidvorstufen. Diese Transporter identifizieren wir einerseits bioinformatisch durch genomweite Suchen mit HMM (hidden Markov model) Profilen, andererseits experimentell mittels phänotypischer Screens. Potentielle Transportergene werden in der Bäckerhefe Saccharomyces cerevisiae exprimiert oder in Oozyten des Krallenfrosches Xenopus laevis. Dank dieser Expressionssysteme lassen sich die physiologischen Substrate und der Transportmechanismus ermitteln. Zudem können wir toxische Substrate identifizieren, welche von den Trypanosomen präferentiell importiert werden. Diese Resultate und Hypothesen testen wir schliesslich in den Trypanosomen selber, wobei uns zu Gute kommt, dass sich diese Parasiten wie kaum andere genetisch manipulieren lassen. So versuchen wir, durch die Kombination verschiedenster Ansätze ein Gesamtbild über die lebenswichtigen Nährstofftransporter von Trypanosoma brucei zu gewinnen und neue Strategien für eine gezielte chemotherapeutische Bekämpfung zu entwickeln.
Direct link to Lay Summary Last update: 27.05.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Arginine and lysine transporters are essential for Trypanosoma brucei
Mathieu C, Macedo JP, Hürlimann D, Wirdnam C, Haindrich AC, Suter Grotemeyer M, Gonzalez-Salgado A, Schmidt RS, Inbar E, Mäser P, Bütikofer P, Zilberstein D, Rentsch D (2017), Arginine and lysine transporters are essential for Trypanosoma brucei, in PLoS One, 12, e0168775.
H+-dependent inorganic phosphate uptake in Trypanosoma brucei is influenced by myo-inositol transporter
Russo-Abrahão T, Koeller CM, Steinmann ME, Silva-Rito S, Marins-Lucena T, Alves-Bezerra M, Lima-Giarola NL, de-Paula IF, Gonzalez-Salgado A, Sigel E, Bütikofer P, Heise N, Gondim KC (2017), H+-dependent inorganic phosphate uptake in Trypanosoma brucei is influenced by myo-inositol transporter, in Meyer-Fernandes, JR, J Bioenerg Biomembr(49), 183.
RFT1 Protein Affects Glycosylphosphatidylinositol (GPI) Anchor Glycosylation
Gottier P, Gonzalez-Salgado A, Menon AK, Liu YC, Acosta-Serrano A, Bütikofer P (2017), RFT1 Protein Affects Glycosylphosphatidylinositol (GPI) Anchor Glycosylation, in J Biol Chem, 292, 1103.
TbIRK is a signature sequence free potassium channel from Trypanosoma brucei locating to acidocalcisomes
Steinmann ME, Schmidt RS, Bütikofer P, Mäser P, Sigel E (2017), TbIRK is a signature sequence free potassium channel from Trypanosoma brucei locating to acidocalcisomes, in Scientific Reports, 7(1), 656.
An Arginine Deprivation Response Pathway Is Induced in Leishmania during Macrophage Invasion
Goldman-Pinkovich A, Balno C, Strasser R, Zeituni-Molad M, Bendelak K, Rentsch D, Ephros M, Jardim A, Wiese M, Myler PJ, Zilberstein D (2016), An Arginine Deprivation Response Pathway Is Induced in Leishmania during Macrophage Invasion, in PLoS Pathogens, 12, e1005494.
An Atypical Mitochondrial Carrier That Mediates Drug Action in Trypanosoma brucei
de Macêdo J (2015), An Atypical Mitochondrial Carrier That Mediates Drug Action in Trypanosoma brucei, in PLoS Pathogens, 11, e1004875.
Flagellar membranes are rich in raft-forming phospholipids
Serricchio M, Schmid AW, Steinmann ME, Sigel E, Rauch M, Julkowska D, Bonnefoy S, Fort C, Bastin P, Bütikofer P (2015), Flagellar membranes are rich in raft-forming phospholipids, in Biol Open, 14, 1143.
Size does matter: 18 amino acids at the N-terminal tip of an amino acid transporter in Leishmania determine substrate specificity
Schlisselberg D, Mazarib E, Inbar E, Rentsch D, Myler PJ, Zilberstein D (2015), Size does matter: 18 amino acids at the N-terminal tip of an amino acid transporter in Leishmania determine substrate specificity, in Scientific Reports, 5, 16289.
TrypanoCyc: a community-led biochemical pathways database for Trypanosoma brucei
Shameer S (2015), TrypanoCyc: a community-led biochemical pathways database for Trypanosoma brucei, in Nucleic Acids Res, 43, D637.
Trypanosoma brucei Bloodstream Forms Depend upon Uptake of myo-Inositol for Golgi Complex Phosphatidylinositol Synthesis and Normal Cell Growth
González-Salgado A (2015), Trypanosoma brucei Bloodstream Forms Depend upon Uptake of myo-Inositol for Golgi Complex Phosphatidylinositol Synthesis and Normal Cell Growth, in Eukaryot Cell, 14, 616.
Trypanosoma brucei eflornithine transporter AAT6 is a low-affinity low-selective transporter for neutral amino acids.
Mathieu Christoph, Salgado Amaia González, Wirdnam Corina, Meier Stefan, Grotemeyer Marianne Suter, Inbar Ehud, Mäser Pascal, Zilberstein Dan, Sigel Erwin, Bütikofer Peter, Rentsch Doris (2014), Trypanosoma brucei eflornithine transporter AAT6 is a low-affinity low-selective transporter for neutral amino acids., in The Biochemical journal, 463(1), 9-18.
A versatile proline/alanine transporter in the unicellular pathogen Leishmania donovani regulates amino acid homoeostasis and osmotic stress responses
Inbar E, Schlisselberg D, Suter Grotemeyer M, Rentsch D, Zilberstein D (2013), A versatile proline/alanine transporter in the unicellular pathogen Leishmania donovani regulates amino acid homoeostasis and osmotic stress responses, in Biochem , 449, 555.
Characterization of choline uptake in Trypanosoma brucei procyclic and bloodstream forms.
Macêdo Juan P, Schmidt Remo S, Mäser Pascal, Rentsch Doris, Vial Henri J, Sigel Erwin, Bütikofer Peter (2013), Characterization of choline uptake in Trypanosoma brucei procyclic and bloodstream forms., in Molecular and biochemical parasitology, 190(1), 16-22.
In silico ionomics segregates parasitic from free-living eukaryotes.
Greganova Eva, Steinmann Michael, Mäser Pascal, Fankhauser Niklaus (2013), In silico ionomics segregates parasitic from free-living eukaryotes., in Genome biology and evolution, 5(10), 1902-9.
A heteromeric potassium channel involved in the modulation of the plasma membrane potential is essential for the survival of African trypanosomes
Steinmann M, A heteromeric potassium channel involved in the modulation of the plasma membrane potential is essential for the survival of African trypanosomes, in FASEB J.

Collaboration

Group / person Country
Types of collaboration
Dr. Philippe Bastin France (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Anant Menon United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Terry Smith Great Britain and Northern Ireland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Jean-Louis Reymond, University of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Dan Zilberstein, Technion Haifa Israel (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Isabel Roditi, University of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Prof. Mike Barrett, University of Glasgow Great Britain and Northern Ireland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Dr. Lucien Rufener, Novartis AH Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure

Scientific events



Self-organised

Title Date Place
Swiss Trypanosomatid Meeting 11.01.2017 Leysin, Switzerland
Drug Resistance: From Mechanisms to Management 10.12.2015 Basel, Switzerland

Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Der Parasit, unser treuer Begleiter? German-speaking Switzerland 2016
Talks/events/exhibitions Spiegelbildliche Moleküle German-speaking Switzerland 2016
Talks/events/exhibitions Medikamente sind selektive Gifte German-speaking Switzerland 2015
Talks/events/exhibitions Pantoffeltier, Amöbe & Co: primitiv oder hochentwickelt? German-speaking Switzerland 2015
Talks/events/exhibitions Parasiten. Life undercover - Führungen German-speaking Switzerland 2014

Awards

Title Year
SNF Early Postdoc Mobility Fellowship 2016

Associated projects

Number Title Start Funding scheme
149229 Transporters for di- and tripeptides in Arabidopsis 01.01.2014 Project funding (Div. I-III)
182426 Amino acid transport, regulation and sensing in Trypanosoma brucei 01.09.2019 Project funding (Div. I-III)
149353 Mitochondrial lipid synthesis in the model eukaryote, Trypanosoma brucei: identification of lipid biosynthesis complexes and contributions of individual pathways to mitochondrial structure and function 01.11.2013 Project funding (Div. I-III)
135746 Drug resistance in African trypanosomes - a deep sequencing approach 01.07.2011 Project funding (Div. I-III)
130815 Novel biosynthetic pathways in Trypanosoma brucei: Biosynthesis of phospholipids and modification of eEF1A 01.08.2010 Project funding (Div. I-III)
127300 Functional genomics of nutrient transporters in Trypanosoma brucei: From physiology to pharmacology 01.01.2010 Sinergia
156264 Drug resistance in African trypanosomes - from the lab to the field and back 01.07.2015 Project funding (Div. I-III)
157884 Supercritical fluid chromatography tandem mass spectrometry as a new analytical tool for plant sciences 01.06.2015 R'EQUIP
156929 Towards an understanding of allosteric modulation of GABA-A receptor isoforms 01.10.2014 Project funding (Div. I-III)

Abstract

Transporters are increasingly being recognized as key players in cell physiology and determinants of drug susceptibility. This is particularly the case for parasites, which - by definition - rely on import of nutrients from their hosts. In our ongoing Sinergia project on functional genomics of nutrient transporters in Trypanosoma brucei, we are investigating four selected groups of nutrient transporters which we believe to be of particular pharmacological interest in the sleeping sickness parasite: ion channels, amino acid transporters, ABC transporters, and transporters of lipid precursors. Substrate specificities and transport kinetics are being determined by expression in Saccharomyces cerevisiae and Xenopus laevis oocytes while the physiological role is studied by reverse genetics in T. brucei. This combination has proven very fruitful. By investigating transporters in an interdisciplinary way, we are validating them as drug targets or drug delivery systems. In the present research proposal we will further extend and deepen these analyses. Incorporating phylogenomic comparisons with fully sequenced trypanosomatid genomes and new developments in the field of T. brucei molecular genetics will allow us to understand the transporters in the context of their phylogeny, physiology and pharmacology.Essentiality of T. brucei nutrient transporters will be predicted in silico based on conserved orthology groups amongst trypanosomatids and imprints of negative selection in the coding regions. Essentiality prediction in vivo will be performed by a combination of genome-wide RNAi screens and high throughput sequencing. Thus nutrient transporters will be prioritized according to their predicted pharmacological and physiological importance. Heterologous expression in S. cerevisiae and X. laevis oocytes functionally characterizes the transporters and serves as a screening platform for inhibitors and toxic substrates of transporters that have been validated as essential in T. brucei bloodstream forms. The subcellular localization of transporters will be determined in trypanosomes, and lipidomics and metabolomics will be applied to study transport physiology and to de-orphanize transporters of unknown substrates. Finally, drug tests against bloodstream-form T. brucei in culture or in a mouse model will validate the identified inhibitors or toxic substrates and identify further leads. Integrated data management will be done on the KNIME (Konstanz Information Miner) platform.The proposed research will result in an increased understanding of the evolution of T. brucei transporters, their physiology in terms of nutrient acquisition and metabolism, and their pharmacological potential for chemotherapeutic strategies that exploit transporters as drug delivery systems or targets. New starting points for drug development against trypanosomatid parasites and potentially novel kinds of nutrient transporters will be discovered.
-