Centromeres mediate the conserved and essential process of chromosome segregation, yet centromeric DNA and the centromeric histone, CENP-A, are rapidly evolving. The rapid evolution of loop 1 (L1) of Drosophila CENP-A is thought to modulate the DNA-binding preferences of CENP-A to suppress centromere drive, the preferential transmission of chromosomes with expanded centromeric satellites during female meiosis. Consistent with this model, CENP-A from D. bipectinata (bip) fails to localize to D. melanogaster (mel) centromeres due to amino acid differences between mel and bip L1. Here, I show that this result is, in fact, due to the inability of the mel CENP-A chaperone, CAL1, to incorporate bip CENP-A into chromatin. Co-expression of bip CENP-A and bip CAL1 in mel cells restores centromeric localization, and similar findings apply to other Drosophila species. Furthermore, two co-evolving regions, CENP-A L1 and the CAL1 N-terminus, are identified as critical for lineage-specific CENP-A incorporation. Collectively, these data show that the rapid evolution of L1 regulates CAL1-mediated CENP-A assembly, suggesting an alternative mechanism for the suppression of centromere drive
