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

我们研究了人着丝粒重复α DNA中着丝粒染色质和/或异染色质组装平衡的机制。我们发现，组蛋白乙酰转移酶（HAT）的alphoid DNA阵列的拴系打破了从头稳定CENP-A染色质组装的细胞类型特异性障碍（异染色质形成），并诱导组装稳定的人类人工染色体（HAC）与输入alphoid DNA。然后，我们将大量tetR融合蛋白与tetO-α HAC连接，并使用KAZUSA全长cDNA文库筛选了许多CENP-A染色质组装的正调控因子和负调控因子。我们还通过融合这些调节因子和CENP-B开发了一个靶向内源性着丝粒的染色质组装平衡的系统。

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.