Analysis of protein-DNA and protein-protein interactions of centromere protein B (CENP-B) and properties of the DNA-CENP-B complex in the cell cycle
Analysis of protein-DNA and protein-protein interactions of centromere protein B (CENP-B) and properties of the DNA-CENP-B complex in the cell cycle
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着丝粒蛋白 B (CENP-B) 的蛋白质-DNA 和蛋白质-蛋白质相互作用分析以及 DNA-CENP-B 复合物在细胞周期中的特性
DOI：
--
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发表时间：
1995
影响因子：
5.3
通讯作者：
Tuneko Okazaki
中科院分区：
文献类型：
--
作者：
Katsumi Kitagawa;Hiroshi Masumoto;Masako Ikeda;Tuneko Okazaki
我们以前报道过着丝粒蛋白B （CENP-B）在体外形成含有两个阿尔法dna的稳定复合物（指定复合物a）。在本研究中，通过大肠杆菌和兔网织细胞裂解物中表达的截断衍生物，确定了参与复合物A形成的CENP-B多肽结构域。凝胶迁移分析表明，含有nh2末端DNA结合结构域的多肽以单体形式结合DNA分子，而在59个氨基酸残基的cooh末端区域的一个新的疏水结构域上二聚。这种多肽在cooh末端的二聚化活性也在酿酒酵母细胞的双杂交体系中得到了证实。结果证明，CENP-B多肽在cooh末端疏水结构域形成同二聚体，每个多肽在其nh2末端结构域结合一条DNA链。二聚化结构域和dna结合结构域位于人和小鼠CENP-B中发现的三个完全保守序列中的两个，并且在小鼠细胞的核提取物中也检测到复合物a形成活性。CENP-B的中期特异性磷酸化也被检测到，但这对其复合物a形成活性没有影响。基于目前的结果，我们提出在间期核和有丝分裂染色体上的着丝粒重复DNA上的CENP-B盒位点形成独特的DNA-蛋白复合物，在着丝粒结构的组装中发挥重要作用。
We previously reported that centromere protein B (CENP-B) forms a stable complex (designated complex A) containing two alphoid DNAs in vitro. Domains in the CENP-B polypeptide involved in the formation of complex A were determined in the present study with truncated derivatives expressed in Escherichia coli and in rabbit reticulocyte lysates. It was revealed by gel mobility shift analyses that polypeptides containing the NH2-terminal DNA-binding domain bind a DNA molecule as a monomer, while dimerizing at a novel hydrophobic domain in the COOH-terminal region of 59 amino acid residues. This polypeptide dimerization activity at the COOH-terminal region was also confirmed with the two-hybrid system in Saccharomyces cerevisiae cells. The results thus proved that CENP-B polypeptides form a homodimer at the COOH-terminal hydrophobic domain, each binding a DNA strand at their NH2-terminal domains. The dimerization and DNA-binding domains fall into two of the three completely conserved sequences found in human and mouse CENP-B, and complex A-forming activity was also detected in nuclear extracts of mouse cells. Metaphase-specific phosphorylation of CENP-B was also detected, but this had no effect on its complex A-forming activity. On the basis of the present results, we propose that CENP-B plays an important role in the assembly of specific centromere structures by forming unique DNA-protein complexes at the sites of CENP-B boxes on the centromeric repetitive DNA both in interphase nuclei and on mitotic chromosomes.
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DOI：
10.1073/pnas.88.9.3734
发表时间：
1991-05-01
影响因子：
11.1
作者：
PALMER, DK;ODAY, K;MARGOLIS, RL
通讯作者：
MARGOLIS, RL
影响因子：
2.9
作者：
GUAN, KL;DIXON, JE
通讯作者：
DIXON, JE
Max 的突变分析：基本螺旋-环-螺旋/亮氨酸拉链结构域在 DNA 结合、二聚化和 Myc 介导的转录激活调节中的作用。
影响因子：
8
作者：
Reddy,CD;Dasgupta,P;Saikumar,P;Dudek,H;Rauscher3rd,FJ;Reddy,EP
通讯作者：
Reddy,EP
影响因子：
56.9
作者：
BLACKWOOD, EM;EISENMAN, RN
通讯作者：
EISENMAN, RN
酵母着丝粒结合蛋白 CP1 的纯化及其结合位点的突变分析。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Baker,RE;Fitzgerald-Hayes,M;O'Brien,TC
通讯作者：
O'Brien,TC