Comparative genomics of drug resistance in Trypanosoma brucei rhodesiense

Fabrice E. Graf, Philipp Ludin, Christian Arquint, Remo S. Schmidt, Nadia Schaub, Christina Kunz Renggli, Jane C. Munday, Jessica Krezdorn, Nicola Baker, David Horn, Oliver Balmer, Adalgisa Caccone (+2 others)
2016 Cellular and Molecular Life Sciences (CMLS)  
Trypanosoma brucei rhodesiense is one of the causative agents of human sleeping sickness, a fatal disease that is transmitted by tsetse flies and restricted to Sub-Saharan Africa. Here we investigate two independent lines of T. b. rhodesiense that have been selected with the drugs melarsoprol and pentamidine over the course of 2 years, until they exhibited stable cross-resistance to an unprecedented degree. We apply comparative genomics and transcriptomics to identify the underlying mutations.
more » ... nly few mutations have become fixed during selection. Three genes were affected by mutations in both lines: the aminopurine transporter AT1, the aquaporin AQP2, and the RNA-binding protein UBP1. The melarsoprol-selected line carried a large deletion including the adenosine transporter gene AT1, whereas the pentamidine-selected line carried a heterozygous point mutation in AT1, G430R, which rendered the transporter non-functional. Both resistant lines had lost AQP2, and both lines carried the same point mutation, R131L, in the RNA-binding motif of UBP1. The finding that concomitant deletion of the known resistance genes AT1 and AQP2 in T. b. brucei failed to phenocopy the high levels of resistance of the T. b. rhodesiense mutants indicated a possible role of UBP1 in melarsoprolpentamidine cross-resistance. However, homozygous in situ expression of UBP1-Leu 131 in T. b. brucei did not affect the sensitivity to melarsoprol or pentamidine. Keywords African trypanosomes Á Aquaporin Á RNA-binding protein Á Purine permease Á Pentamidine Á Melarsoprol Abbreviations MPXR Melarsoprol-pentamidine cross-resistance SLT Spliced leader trapping AQP2 Aquaglyceroporin 2 AT1 Adenosine transporter 1 Tet Tetracycline UBP1 Uridine-rich binding protein 1 VSG Variant surface glycoprotein
doi:10.1007/s00018-016-2173-6 pmid:26973180 pmcid:PMC4967103 fatcat:pcl62svulvbv3jvkfquv5ne2c4