Enrichment and mass spectrometry analysis of TbTAX-1-interacting or TbTTC29-interacting proteins by co-immunoprecipitation

Study of Trypanosoma ZMYND12’s ortholog (TbTAX-1) confirmed an axonemal localization of the protein and knockdown of TbTAX-1 using RNAi induced dramatic flagellar motility defects as observed in human. Co-immunoprecipitation assays and ultra-structure expansion microscopy in Trypanosoma brucei then formally demonstrated that ZMYND12 and TTC29 directly interact and belong to the same axonemal complex. Subsequent comparative proteomics analysis in Trypanosoma and Ttc29 KO mice models also identified DNAH1 as a new interacting protein of both ZMYND12 and TTC29 proteins. These data evidenced the critical role of ZMYND12 for flagellum function and assembly in humans and Trypanosoma through its interaction with other axonemal partners such as TTC29 and DNAH1 in the same complex.