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Analysis of functienal domain and expression in bone, cartilage and muscle of minor collagen

小胶原蛋白在骨、软骨和肌肉中的功能域和表达分析

基本信息

批准号：
17390418
负责人：
YOSHIOKA Hidekatsu
金额：
$ 10.56万
依托单位：
Oita University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

1) Initially, to determine the transcription initiation site of mouse α3 (V) gene, Oligo-Cap methods was used. It was located about 100 bp upstream of ATG codon. We used structure-function analysis of the core promoter region to elucidate the transcriptional features of the gene. The core promoter was defined within about 300 bp immediately upstream from the transcription start site by transient transfection experiments. In this region, we identified two nuclear-factor binding sites by electrophoretic mobility shift assay (EMSAs): BS1 (-130 to -110), BS2 (-190 to -170). Oligonucleotide competition and supershift assays revealed that CBF,/NF-Y specifically bind to BS1.The N-terminal globular domain of the pro-α3 (V) chain has a unique structure that contains a highly basic segment of 23 amino acids. The peptide showed a specific adhesive feature to osteosarcoma cells but not to chondrosarcoma cells. This feature was derived from four heparin binding sites in the segment. When the peptid … More e adhere to the cell, actin fiber was formed, which was inhibited Rho-kinase inhibitor, Y27632.2) We have characterized the proximal promoter of the human α1 (III) collagen gene (COL3A1). Transient transfection assays indicated that the segment from -96 to -34 was necessary to activate transcription. EMSAs showed that the multiple proteins form the DNA-protein complex in different combinations depending on the cell types. The region of -79 to -63 bp was critical for DNA protein complex formation. This sequence was contained in the B element of mouse al (III) collagen gene (Col3a1) reported by other researcher.3) Collagen XXIV is a newly discovered and poorly characterized member of the fibril-forming family of collagen molecules, which is expressed predominantly in bone tissue. Cell transfection experiments in combination with DNA binding assay demonstrated that Co124a1 promoter activity was under control of an upstream cis-acting element, which was recognized by specific combination of c jun, CREB1, ATF1, and ATF2 dimers. Less

1)初步采用Oligo-Cap法确定小鼠α3 (V)基因的转录起始位点。它位于ATG密码子上游约100bp处。我们利用核心启动子区域的结构-功能分析来阐明该基因的转录特征。通过瞬时转染实验，在转录起始位点上游约300 bp处确定了核心启动子。在这一区域，我们通过电泳迁移位移测定（EMSAs）确定了两个核因子结合位点：BS1(-130至-110),BS2（-190至-170）。寡核苷酸竞争和超移位实验显示CBF，/NF-Y特异性结合BS1。前α3 (V)链的n端球形结构域具有独特的结构，包含23个氨基酸的高碱性片段。该肽对骨肉瘤细胞有特异的粘附作用，但对软骨肉瘤细胞无粘附作用。这一特征来源于片段中的四个肝素结合位点。当肽更多地粘附于细胞时，形成肌动蛋白纤维，抑制rho激酶抑制剂Y27632.2)。我们已经鉴定了人α1 （III）胶原基因的近端启动子（COL3A1）。瞬时转染实验表明-96到-34段是激活转录所必需的。emsa显示，多种蛋白质根据细胞类型以不同的组合形式形成dna -蛋白质复合物。-79 ~ -63 bp区域是DNA蛋白复合物形成的关键区域。该序列包含在其他研究者报道的小鼠al （III）胶原蛋白基因（Col3a1）的B元件中。3)胶原蛋白XXIV是新发现的纤维形成胶原分子家族的成员，其主要表达于骨组织中。细胞转染实验和DNA结合实验表明，Co124a1启动子活性受上游顺式作用元件控制，该元件通过c jun、CREB1、ATF1和ATF2二聚体的特异性组合识别。少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Multifactor complex containing B element binding factor, BBF, and repressors regulate the human α1(III) collagen gene(COL3A1)

含有 B 元素结合因子、BBF 和阻遏物的多因子复合物调节人类 α1(III) 胶原蛋白基因 (COL3A1)

DOI：
发表时间：
2006
期刊：
Acta Med. Okayama 60
影响因子：
0
作者：
Sumiyoshi;H.;Matsuo;N.;Shin;T.;Shirabe;K.;Yoshioka;H
通讯作者：
H

Adiponectin is expressed inthebrown adipose tissue and surrounding immature tissues in mouse embryo.

脂联素在小鼠胚胎的棕色脂肪组织和周围未成熟组织中表达。

DOI：
发表时间：
2005
期刊：
Biochemica et Biophysica Acta 1731・1
影响因子：
0
作者：
Fujimoto T;Matsuo T;Sumiyoshi H;Yamaguchi K;Saikawa T;Yoshimnatsu H;Yoshioka H
通讯作者：
Yoshioka H

Association between heat stress protein 70 induction and decreased pulmonary fibrosis in an animal model of acute lung injury