Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of NDC80

Protein (function)
NDC80 is a kinetochore marker. Chromosome attachment to spindle microtubules (MTs) is mediated by kinetochores, which are large multiprotein complexes assembled on centromeres located at the constriction point of sister chromatids. Each sister chromatid has its own kinetochore, oriented to facilitate movement to opposite poles of the spindle apparatus. During anaphase, the spindle elongates and the sister chromatids separate, resulting in segregation of the two genomes during telophase. The NDC80 complex is the major component of the kinetochore and mediates its attachment to spindle MTs. In most model organisms, it is a member of the network of conserved KNL1, MIS12 and NDC80 complexes (KMN)
The ∼170–190 kDa NDC80 complex has two globular domains at either end of a ∼57 nm elongated coiled-coil, forming a dumb-bell shape. It is a heterotetramer comprising a 1:1:1:1 ratio of NDC80 (also known as HEC1 in humans), NUF2, SPC24 and SPC25 sub-complexed as two heterodimers: NDC80 with NUF2, and SPC24 with SPC25

Phenotype
the spatiotemporal profile of NDC80–GFP protein expression and location was examined during the parasite life cycle at the three asexual mitotic replicative stages: liver and blood schizogony in the vertebrate host and oocyst development (sporogony) in the mosquito vector, the sexual mitotic stage (male gametogenesis) and the meiotic stage (ookinete development). 

Evidence is presented that:

- Real-time live-cell imaging using NDC80–GFP reveals kinetochores aggregate as discrete foci during schizogony
- Immunofluorescence imaging shows the arrangement of NDC80 (kinetochore) with centrin (putative MTOC/SPB) and α-tubulin (spindles)
- Observation of NDC80–GFP spindle dynamics reveals unusual kinetochore bridges during endoreduplication in male gametogenesis
- NDC80–GFP shows unusual dynamics throughout the meiotic stages during zygote to ookinete differentiation
- NDC80–GFP is present as multiple foci during oocyst development and sporozoite formation

- Immunoprecipitation of NDC80–GFP recovers canonical members of the NDC80 complex and reveals a highly divergent SPC24-like candidate: PBANKA_1442300, a protein with a long coiled-coil region and a predicted C-terminal globular domain, which suggested that it might be a SPC24 orthologue

Additional information

Other mutants