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© Christine Schmitt, Sophie Goyard, Jean-Marc Panaud
Trypanosoma cruzi - trypomastigote form. Trypanosoma cruzi is the etiological agent of Chagas disease.

About

The main focus of the group is to understand the mechanisms of antigenic variation and DNA repair in African trypanosomes, two processes that are intimately linked and essential for virulence. Trypanosoma brucei is a ubiquitous parasite that causes Human African Trypanosomiasis (HAT) in the human host and Nagana in cattle. The broad range of hosts that T. brucei is able to infect make both the medical burden and its economic impact, in terms of daily adjusted life years and rendering large tracts of land unusable for agriculture, devastating to the regions where infection is endemic. Although current treatments suffer from high toxicity and increasing resistance, without treatment HAT is fatal. Trypanosomes are transmitted from the tsetse fly, the insect host, to the mammalian host where they exist and divide in the bloodstream and tissue fluid.

ImageJ=1.48s
Bloodstream form T. brucei cell cycle. VSG, red; DNA, white.

Trypanosomes are covered in approximately 10 million molecules of a single variant surface glycoprotein (VSG), which forms a highly immunogenic surface coat. It is against this  coat that the host’s immune response is directed. Antigenic variation is the stochastic switching of the VSG coat, and allows the trypanosome to evade the host’s adaptive immunity. Fundamental to antigenic variation is strict monoallelic expression of a single  VSG gene and the concomitant silencing of all others. To date, little is  known about how monoallelic expression is controlled in the African trypanosome or how the cells switch  VSG`s .

Picture1
T. brucei expressing either VSG-6, green or VSG-2, red; DNA, white.

Trypanosomes have evolved to usurp DNA recombination as an essential component of VSG switching. VSG gene conversion events that replace the expressed VSG with one from the VSG archive dominate in T. brucei. These events are dependent on homologous recombination, so in order to better understand antigenic variation we need to better understand DNA recombination and repair. Using established genetic tools we can track DNA repair in T. brucei using DNA damage response markers such as RAD51 and phosphorylated histone H2A. Although homologous recombination appears to be conserved in T. brucei, some components show significant sequence diversity, suggesting functional diversity, which may evolved from the dependence on recombination pathways for antigenic variation.

Picture1
RAD51 and gamma H2A mark DNA damage in T. brucei.

Using RNA Interference Target screens coupled with NGS (RIT-seq), molecular, biochemical and cell-biological approaches, in high-throughput mode where appropriate, we aim to tackle these longstanding questions that were intractable using prior technologies.

Former Members

2000
2000
Name
Position
2016
2019
Ross P. Coron
Post-doc
2016
2019
Alix Thivolle
Graduate Student
2018
2019
Eliane Tihon
Post-doc
2017
2021
Emilia MacLaughlin
Doctorant PPU
2018
2021
Nuria Sima Teruel
Post-doc

Projects

Transversal Project

Fundings

Publications

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Lab news

May 2019: Ann-Kathrin has finished her M2 internship in the lab, and will return to Heidelberg to complete her Master’s. She will be very missed in the lab! April 2019: The lab has been to the Kinetoplast Molecular Cell Biology meeting in Woods Hole, Massachusetts where Eliane, Nuria and Emilia each presented a poster. A great time was had by all and we have returned with lots of new research ideas! January 2019: Masters 2 student Ann-Kathrin Mehnert has joined the lab for a four month internship. Ann-Kathrin is completing a masters in Infectious Diseases at the University of Heidelberg, and during her internship will investigate the role of Mre11 in DNA repair in T.brucei. November 2018: CRISPR in parasitology symposium, Institut Pasteur. The TMB lab attended this fantastic symposium, with a specific focus on trouble shooting and sharing tips and ideas to overcome hurdles associated with using CRISPR in parasitology. Nuria gave a talk entitled ‘Multipurpose dCas9 in Trypanosoma brucei’. September 2018: The lab has attended the EMBO workshop ‘Molecular advances and parasites strategies in host infection’ in Les Embiez Island. The meeting was a great opportunity to share ideas with other parasitologists and enjoy some late summer sun! July 2018: Alix has graduated from her master’s. Alix worked in the lab for a total of nine months on her master’s project investigation position dependent repair in the active expression site. She will be greatly missed in the lab! May 2018: Emilia and Lucy have been to the Paris-Pasteur University PhD program retreat in Le Tourquet. Emilia presented a poster entitled ‘DNA Damage in African Trypanosomes’ and won the prize for the best poster teaser! April 2018: The whole lab has attended the Parasites and Insect Vector annual departmental retreat at Château du Maintenon in the Loire Valley. Alix, Nuria, Emilia and Eliane all presented posters, with Alix winning the poster competition for her poster entitled ‘Position dependent repair in the active expression site’. A great time was had by all! February 2018: A second post-doc, Nuria Sima Teruel has started in the lab. Nuria will be looking at the proteins involved in DNA repair in trypanosomes. February 2018: Emilia has completed her first year thesis advisory committee meeting. January 2018: A new post-doc, Eliane Thon, has arrived in the lab. Eliane has received an AAPG-ANR grant and will be working on procyclic and metacylic form parasites. November 2017: Annick has recieved the ‘Mèdail or d’honneur du travail’ for 35 years of service at Institut Pasteur! She began working here in 1982 in a molecular biology laboratory and has worked in seven labs at the Institut.

Contact

Phone: +33 (0)1 40 61 34 25 Email: lucy.glover@pasteur.fr Address 25-28 Rue du Docteur Roux 75015, Paris France Follow us on twitter @Glover_Tryplab