CENP-A is a centromere-specific histone 3 variant essential for centromere specification. CENP-A partially replaces canonical histone H3 at the centromeres. How the particular CENP-A/H3 ratio at centromeres is precisely maintained is unknown. It also remains unclear how CENP-A is excluded from non-centromeric chromatin. Here, we identify Ccp1, an uncharacterized NAP family protein in fission yeast that antagonizes CENP-A loading at both centromeric and non-centromeric regions. Like the CENP-A loading factor HJURP, Ccp1 interacts with CENP-A and is recruited to centromeres at the end of mitosis in a Mis16-dependent manner. These data indicate that factors with opposing CENP-A loading activities are recruited to centromeres. Furthermore, Ccp1 also cooperates with H2A.Z to evict CENP-A assembled in euchromatin. Structural analyses indicate that Ccp1 forms a homodimer that is required for its anti-CENP-A loading activity. Our study establishes mechanisms for maintenance of CENP-A homeostasis at centromeres and the prevention of ectopic assembly of centromeres. Display omitted •Ccp1 antagonizes CENP-A loading at both centromeric and non-centromeric regions•Ccp1 is recruited to centromeres at the end of mitosis in a Mis16-dependent manner•Ccp1 cooperates with H2A.Z to evict CENP-A assembled in euchromatin•Ccp1 forms a homodimer that is required for its anti-CENP-A loading activity Dong et al. identify Ccp1, a NAP family protein, antagonizing the loading of CENP-A at both centromeric and non-centromeric regions. This study provides insights into both how the balance of CENP-A and histone H3 levels is achieved at centromeres and how non-centromeric regions are protected from mistakenly assembling CENP-A.