The Plasmodium falciparum centromeric histone variant PfCENH3 has been shown to occupy a 4-4.5 kb region on each chromosome, but the experimental demonstration of its structure-function relationship remains unexplored. By functional complementation assays, we report that the C-terminus, specifically the CATD region within the HFD of PfCENH3 is essential in centromere function. Our studies also indicate that the PfCENH3 specific LLAL residues of the CATD region are required for centromere targeting and chromosome segregation. Histone H3 of P. falciparum is not found to complement Cse4p (the yeast homologue of CENH3). We also report the identification of PfCENP-C, another component of the inner kinetochore protein complex and its association with PfCENH3. These studies thus delineate the structural determinants of PfCENH3.
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