Analysis of Intranuclear Communication of Homologous Alleles of Imprinted Genes and Transgenes

印记基因和转基因同源等位基因的核内通讯分析

基本信息

批准号：
11660086
负责人：
OKUMURA Katsuzumi
金额：
$ 2.3万
依托单位：
Mie University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11660086/
关键词：
Genomic imprinting FISH DNA methylation Histone acetylation Nuclear matrix Heterochromatin SNRPN 一次転写産物ヒストン脱アセチル化 genome imprinting intranuclear structure Igf2r Chromosome territory gene silencing heterochromatin

项目摘要

Investigations of imprinted regions provide clues that increase our understanding of the regulation of gene functions at higher order chromosomal domains. The relative positions of the chromosome 15 centromere and the imprinted SNRPN gene in interphase nuclei of human myeloid leukemia HL60 cells were compared by multicolor FISH, because the homologous association of this imprinted chromosomal domain was previously observed in lymphocytes and lymphoblasts. Preferential association of SNRPN interhomologues does not occur during the cell cycle, although this gene exhibited asynchronous replication and monoallelic expression. SNRPN was found to localize at the periphery of the chromosome territories, and it preferentially faces the nuclear membrane. The SNRPN gene and the centromere are located close to each other late in S phase, reflecting that these DNA segments may be compacted into the same intranuclear subcompartments with the progress of S phase. Our results suggest that the charact … More eristic of mutual recognition of imprinted regions is determined by certain cellular regulation, and it is not necessary for the allele-specific features of an imprinted gene. We have adapted a FISH method to detect nascent RNA molecules of the imprinted SNRPN gene at the initial transcription sites in nuclei of human HL60 and WI38 cells. Simultaneous detection of RNA and DNA of SNRPN by FISH using the cosmid probe confirmed its monoallelic expression in these cell lines. Treatment of both cells by inhibitors of DNA methylation and histone deacetylation resulted in time-dependent increase of the cell population with the biallelic expression of SNRPN.We investigated the allele-specific gene silencing of imprinted regions by their association with heterochromatin. Our results suggested the possibility and dependency on DNA replication. We further examined the relationship between gene expression and nuclear matrix by using imprinted genes, suggesting that the expressed allele associated with the nuclear matrix. Less

对印迹区域的研究提供了线索，以增加我们对高阶染色体域基因功能调节的理解。通过多色鱼类比较了人髓样白血病HL60细胞中染色体15共粒和印迹SNRPN基因的相对位置。尽管该基因暴露了异步复制和单相表达，但在细胞周期期间并未发生SNRPN跨学科的优先关联。发现SNRPN位于染色体区域的外围，它优先面对核膜。 SNRPN基因和中心仪在S期的后期彼此靠近，这反映了这些DNA片段可以随着S相进展而将这些DNA片段压缩到相同的内部分析子组件中。我们的结果表明，特征……对烙印区域的相互认识更加敏锐，是由某些细胞调节决定的，并且对于烙印基因的等位基因特定特征并不是必需的。我们已经改编了一种鱼方法，以检测核HL60和Wi38细胞的初始转录位点上印迹SNRPN基因的新生RNA分子。使用宇宙探针同时检测鱼类的RNA和DNA，证实了其在这些细胞系中的单相表达。通过DNA甲基化和组蛋白脱乙酰化的抑制剂对两种细胞进行处理，导致细胞群的时间依赖性增加，而SNRPN的生物素表达进行了研究。我们研究了等位基因特异性的基因沉默的印记区域通过与异染色质的关联。我们的结果表明了对DNA复制的可能性和依赖性。我们通过使用印迹基因进一步研究了基因表达与核基质之间的关系，这表明表达等位基因与核基质相关。较少的