Dynamic chromatin changes associated with de novo centromere formation in maize euchromatin

The inheritance and function of centromeres are not strictly dependent on any specific DNA sequence, but involve an epigenetic component in most species. CENH3, a centromere histone H3 variant, is one of the best-described epigenetic factors in centromere identity, but the chromatin features required during centromere formation have not yet been revealed. We previously identified two de novo centromeres on Zea mays (maize) minichromosomes derived from euchromatic sites with high-density gene distributions but low-density transposon distributions. The distribution of gene location and gene expression in these sites indicates that transcriptionally active regions can initiate de novo centromere formation, and CENH3 seeding shows a preference for gene-free regions or regions with no gene expression. The locations of the expressed genes detected were at relatively hypomethylated loci, and the altered gene expression resulted from de novo centromere formation, but not from the additional copy of the minichromosome. The initial overall DNA methylation level of the two de novo regions was at a low level, but increased substantially to that of native centromeres after centromere formation. These results illustrate the dynamic chromatin changes during euchromatin-originated de novo centromere formation, which provides insight into the mechanism of de novo centromere formation and regulation of subsequent consequences. Significance Statement Centromeres are often embedded in heterochromatin-rich and transcriptionally inert regions. Here we investigated two maize de novo centromeres atypically located in euchromatin. The dynamic changes of DNA methylation patterns indicate that seeding of CENH3, the centromere-specific histone, was affected by intrinsic DNA methylation patterns before neocentromere formation and that CENH3 loading can also shape the DNA methylation patterns after de novo centromere formation.