Chromatin assembly factor-1 (CAF-1) is essential for chromatin assembly in eukaryotes, and comprises three subunits of 48 kDa (p48), 60 kDa (p60), and 150 kDa (p150). The small subunit of the chicken CAF-1, chCAF-1p48, possesses seven WD repeat motifs, contains only one amino acid change relative to the human and mouse CAF-1p48s, and interacts with chicken histone deacetylases (chHDAC-1 and 2) in vivo. The GST pulldown affinity assay revealed the in vitro interaction of chCAF-1p48 with chHDAC-1, 2 and 3. We established that the p48 subunit tightly binds to two regions of chHDAC-2 comprising amino acid residues 82-180 and 245-314, respectively. In addition, two N-terminal, two C-terminal, or one N-terminal and one C-terminal WD repeat motif of chCAF-1p48 are required for this interaction. These result suggest that chCAF-1p48 is involved in many aspects of DNA-utilizing processes, through alterations in the chromatin structure based on both the acetylation and deacetylation of core histones.