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CHD1 is a SNF2-related ATPase that is required for the genome-wide incorporation of variant histone H3.3 in the paternal pronucleus as well as in transcriptionally active nuclei in  Drosophila  embryos. The  S. pombe  and vertebrate orthologs of CHD1 have been implicated in the assembly of the centromeric histone H3 variant CenH3 CENP-A , which occurs in a DNA replication-independent manner. Here, we examined whether CHD1 participates in the assembly of CenH3 CID  in  Drosophila . In contrast to the findings in fission yeast and vertebrate cells, our evidence clearly argues against such a role for CHD1 in  Drosophila . CHD1 does not localize to centromeres in either S2 cells or developing fly embryos. Down-regulation of CHD1 in S2 cells by RNAi reveals unchanged levels of CenH3 CID  at the centromeres. Most notably, ablation of functional CHD1 in  Chd1  mutant fly embryos does not interfere with centromere and kinetochore assembly, as the levels and localization of CenH3 CID , CENP-C and BubR1 in the mutant embryos remain similar to those seen in wild-type embryos. These results indicate that  Drosophila  CHD1 has no direct function in the incorporation of the centromeric H3 variant CenH3 CID  into chromatin. Therefore, centromeric chromatin assembly may involve different mechanisms in different organisms.

CenH3/CID Incorporation Is Not Dependent on the Chromatin Assembly Factor CHD1 in Drosophila