The kinesin of the flagellum attachment zone in Leishmania is required for cell morphogenesis, cell division and virulence in the mammalian host
Abstract
Leishmania parasites possess a unique and complex cytoskeletal structure termed flagellum attachment zone (FAZ) connecting the base of the flagellum to one side of the flagellar pocket (FP), an invagination of the cell body membrane and the sole site for endocytosis and exocytosis. This structure is involved in FP architecture and cell morphogenesis, but its precise role and molecular composition remain enigmatic.
Here, we characterized Leishmania FAZ7, the only known FAZ protein containing a kinesin motor domain, and part of a clade of trypanosomatid-specific kinesins with unknown functions. The two paralogs of FAZ7, FAZ7A and FAZ7B, display different localizations and functions.
FAZ7A localizes at the basal body, while FAZ7B localizes at the distal part of the FP, where the FAZ structure is present in Leishmania. While null mutants of FAZ7A displayed normal growth rates, the deletion of FAZ7B impaired cell growth in both promastigotes and amastigotes of Leishmania.
The kinesin activity is crucial for its function. Deletion of FAZ7B resulted in altered cell division, cell morphogenesis (including flagellum length), and FP structure and function.
Furthermore, knocking out FAZ7B induced a mis-localization of two of the FAZ proteins, and disrupted the molecular organization of the FP collar, affecting the localization of its components. Loss of the kinesin FAZ7B has important consequences in the insect vector and mammalian host by reducing proliferation in the sand fly and pathogenicity in mice.
Our findings reveal the pivotal role of the only FAZ kinesin as part of the factors important for a successful life cycle of Leishmania. Author summary Leishmania are flagellated trypanosomatid parasites causing worldwide human and animal diseases.
As 'divergent eukaryotes', their biology presents unique features and structures, of which the specific functions constitute potential drug targets. Among others, they possess a unique cytoskeletal structure termed the flagellum attachment zone (FAZ) attaching the base of their flagellum to one side of the flagellar pocket (FP), which is the sole site for endocytosis and exocytosis.
The FP together with other unique flagellum-associated structures are crucial for parasite survival, but the functioning of this whole remains largely enigmatic. Leishmania also possess an expanded repertoire of kinesins (>55), including two trypanosomatid-specific families.
Here, we show that the deletion of the sole kinesin among FAZ proteins disrupts cell morphogenesis, FP organisation and cell division. Furthermore, the ability to proliferate in the insect vector and mammalian host is reduced in parasites lacking the kinesin FAZ7B.
This study helps elucidate the factors contributing to the successful lifecycle and pathogenicity of the parasite. It also highlights the functional diversification of motor proteins during evolution.