Taxa-specific proteins are key determinants defining the biology of all organisms and rep-resent prime drug targets in pathogens. How-ever, lacking comparability with proteins in other lineages makes them particularly diffi-cult to study. In malaria parasites this is exac-erbated by technical limitations. Here, we ana-lysed the cellular location, essentiality, func-tion and, in selected cases, interactome of all unknown non-secretory proteins encoded on an entire P. falciparum chromosome. The nu-cleus was the most common localisation, in-dicating it is a hotspot of parasite-specific bi-ology. More in-depth functional studies with four proteins revealed essential roles in DNA replication and mitosis. The novel mitosis pro-teins defined a possible orphan complex and a highly diverged complex needed for the spin-dle-kinetochore connection. Structure-function comparisons indicated that the taxa-specific proteins evolved by different mecha-nisms. This work demonstrates the feasibility of gene-by-gene screens to elucidate the biol-ogy of malaria parasites and reveal critical parasite-specific processes of interest as drug targets.
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