Identification of a basal body-localized epsilon-tubulin in Leishmania

Abstract

Epsilon-tubulins have been identified in only a few organisms to date. We identified an epsilon-tubulin in Leishmania major (Lmε-tubulin) exhibiting significant sequence similarity and conservation of functional domains with its counterparts. It is expressed in both promastigote and amastigote forms of the parasite. For localization studies, we generated a Leishmania strain expressing the endogenous Lmε-tubulin tagged with mNeonGreen by CRISPR-Cas9. Imaging of this strain revealed that Lmε-tubulin is located near the kinetoplast and at the flagellar base, indicating basal body localization. Lmε-tubulin is not part of the microtubular network since its localization remained unaltered upon treatment with nocodazole, a microtubule disruptor. This is the first experimental identification of an ε-tubulin not only in Leishmania but in the entire Trypanosomatidae family.