Abstract
During the Leishmania life cycle, the flagellum undergoes successive assembly and disassembly of hundreds of proteins. Understanding these processes necessitates the study of individual components. Here, we investigated LdFlabarin, an uncharacterized L. donovani flagellar protein. The gene is conserved within the Leishmania genus and orthologous genes only exist in the Trypanosoma genus. LdFlabarin associates with the flagellar plasma membrane, extending from the base to the tip of the flagellum as a helicoidal structure. Site-directed mutagenesis, deletions and chimera constructs showed that LdFlabarin flagellar addressing necessitates three determinants: an N-terminal potential acylation site and a central BAR domain for membrane targeting and the C-terminal domain for flagellar specificity. In vitro, the protein spontaneously associates with liposomes, triggering tubule formation, which suggests a structural/morphogenetic function. LdFlabarin is the first characterized Leishmania BAR domain protein, and the first flagellum-specific BAR domain protein.
Highlights
Eukaryotic flagella present a remarkable evolutionary conservation of their structure and constituents [1]
Identification of the Leishmania Flabarin We previously discovered the involvement of small G proteins in the biogenesis of the Leishmania flagellum [21,22,23], which led us to try and identify effectors
We report the characterization of Flabarin, a novel Leishmania flagellar protein
Summary
Eukaryotic flagella present a remarkable evolutionary conservation of their structure and constituents [1] They have been mostly studied in Chlamydomonas reinhardtii [2,3] and Trypanosoma brucei [4,5], two flagellated organisms. Intraflagellar transport was first described in C. reinhardtii before being generalized to all eukaryotic flagella and cilia [6,7] Stressing their practical importance, protozoan studies allowed the identification of human orthologue genes whose mutations are responsible for pleiotropic, severe genetic diseases, such as polycystic kidney disease or BardetBiedl syndrome [8]. Leishmania are flagellated protozoan kinetoplastid parasites [9] They exist alternatively as amastigotes (intracellular mammalian forms) and promastigotes (extracellular insect forms) [10]. The dynamics of these processes is tightly regulated and their understanding necessitates the characterization of individual elements
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