Functional Structure of Centromere/Kinetochore Domain of Mammalian Chromosomes

哺乳动物染色体着丝粒/动粒结构域的功能结构

• 批准号: 01480537
• 负责人: OKAZAKI Tuneko
• 金额: $ 4.42万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480537/
• 关键词: Antigenic protein recognized by ACA; CENP-B; alphoid DNA; CENP-B box; DNA-protein interaction; centromere; kinetochore; 17塩基対モチ-フ; DNAー蛋白相互作用; CENP-B; DNA-蛋白相互作用

The purpose of this research is to obtain the basic information reguired to understand the regulation mechanism of chromosome segregation through the identification of DNA sequence and protein factors responsible for the functional structure of centromere/kinetochore of mammalian chromosomes. Following information was obtained.1. Highly repetitive alpha-satellite DNA (alphoid DNA) is localized at centromere domain.2. Three antigens {CENP-A (17 kD), CENP-B (80 kD) and CENP-C (140 kD)} are detected with anticentromere antibodies in sera from autoimmune patients. These antigens are specifically associated with centromere domain of human chromosomes.3. CENP-B and alphoid DNA form a complex at the 17 bp motif (CENP-B box) in alphoid DNA.4. CENP-B box sequences are detected at centromere domain of all human chromosomes, including Y-chromosome in which only very small amount seems to be present. The CENP-B box is also found in centromeric satellite DNA of mouse.5. Centromere antigens which form a complex with the CENP-B box are found in nuclear extracts of mouse, Indian muntjac and monkey cells.6. A complex formed between alphoid DNA and CENP-B consists of two DNA molecules and probably two CENP-B molecules.Thus, CENP-B/CENP-B box interaction is widely found in mammalian chromosomes and this interaction seems to play a role in higher order formation of centromeric alphoid DNA.

本研究的目的是通过对哺乳动物染色体着丝粒/动粒功能结构的DNA序列和蛋白质因子的鉴定，为了解染色体分离的调控机制提供基础信息。获得以下信息。1.高度重复的α-卫星DNA（alphoid DNA）定位于着丝粒结构域.用抗着丝粒抗体在自身免疫患者血清中检测到三种抗原{CENP-A（17 kD）、CENP-B（80 kD）和CENP-C（140 kD）}。这些抗原与人类染色体的着丝粒区特异性结合. CENP-B与α型DNA在α型DNA中的17 bp基序（CENP-B box）处形成复合物。CENP-B盒序列在所有人类染色体的着丝粒结构域处检测到，包括Y染色体，其中似乎仅存在非常少量。CENP-B盒也存在于小鼠的着丝粒卫星DNA中.与CENP-B盒形成复合物的着丝粒抗原存在于小鼠、印度山羌和猴细胞的核提取物中。α DNA与CENP-B形成的复合物由两个DNA分子和可能的两个CENP-B分子组成，因此，CENP-B/CENP-B box相互作用在哺乳动物染色体中广泛存在，这种相互作用可能在着丝粒α DNA的高级形成中起作用。

项目成果

Hiroshi MAsumoto: "A human centromere antigen (CENPーB) interacts with a short specific sequence in alphoid DNA,a human centromric satellite." J.Cell Biol.109. 1963-1973 (1989)

Hiroshi MAsumoto：“人类着丝粒抗原 (CENP-B) 与人类着丝粒卫星中的短特定序列相互作用。”J.Cell Biol.109 (1989)。

Yoshinao Muro: "The heterogeneity of anticentromere antibodies in immunoblotting analysis." J.Rheumatology. 17. 1042-1047 (1990)

Yoshinao Muro：“免疫印迹分析中抗着丝粒抗体的异质性。”

Hiroshi Masumoto :"Alphoid satellite DNA is tightly associated with centromere antigens in human chromosomes throughout the cell cycle." Exp. Cell Res.181. 181-196 ((198)

Hiroshi Masumoto ：“在整个细胞周期中，Alphoid 卫星 DNA 与人类染色体中的着丝粒抗原紧密相关。”

H.Masumoto: "Alphoid satellite DNA is tightly associated with centromere antigens in human chromosomes throughout the cell cycle." Exp.Cell Res.181. 181-196 (1989)

H.Masumoto：“在整个细胞周期中，Alphoid 卫星 DNA 与人类染色体中的着丝粒抗原紧密相关。”

Hiroshi Masumoto: "Alphoid Satellite DNA is tightly associated with centromere antigens in human chromosomes throughout the cell cysle." Exp.Cell Res.181. 181-196 (1989)

Hiroshi Masumoto：“Alphoid Satellite DNA 与整个细胞周期中人类染色体的着丝粒抗原紧密相关。”