Analysis of mechanism involved in the assembly balance between centromere chromatin and/or heterochromatin and in higher cellular regulation.

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We investigated mechanism involved in the chromatin assembly balance between centromere chromatin and/or heterochromatin at human centromeric repetitive alphoid DNA. We found that tethering of histone acetyltransferases (HATs) to alphoid DNA arrays broke a cell type-specific barrier (heterochromatin formation) for de novo stable CENP-A chromatin assembly and induced assembly of stable Human Artificial Chromosomes (HACs) with the input alphoid DNA. We, then, tethered a huge variety of tetR-fusion proteins to the tetO-alphoid HAC and screened many positive and negative regulators for the CENP-A chromatin assembly using KAZUSA full-length cDNA library. We also developed a system targeting the chromatin assembly balance at endogenous centromeres by fusing these regulators and CENP-B.