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

印记区域的研究提供了增加我们对高阶染色体区域基因功能调控的理解的线索。通过多色 FISH 比较了人骨髓性白血病 HL60 细胞间期细胞核中 15 号染色体着丝粒和印记 SNRPN 基因的相对位置。因为先前在淋巴细胞和淋巴母细胞中观察到该印记染色体结构域的同源关联。尽管该基因呈现异步复制并且单等位基因表达位于染色体区域的外围，并且它优先面向核膜和着丝粒，但在细胞周期中不发生同源同源。彼此在 S 期晚期相互结合，反映出随着 S 期的进展，这些 DNA 片段可能被压缩到相同的核内亚区室中。印记区域的相互识别是由某些细胞调节决定的，对于印记基因的等位基因特异性特征来说，这不是必需的。我们采用了 FISH 方法来检测印记 SNRPN 基因在初始转录位点的新生 RNA 分子。使用粘粒探针对人 HL60 和 WI38 细胞的细胞核中 SNRPN 的 RNA 和 DNA 进行同时检测，证实了这两种细胞系中 SNRPN 的单等位基因表达。 DNA 甲基化和组蛋白脱乙酰化导致细胞群随 SNRPN 双等位基因表达而呈时间依赖性增加。我们通过印记区域与异染色质的关联研究了等位基因特异性基因沉默，我们的结果表明了 DNA 复制的可能性和依赖性。我们通过使用印记基因进一步检查了基因表达与核基质之间的关系，表明表达的等位基因与核基质相关较少。