Exploration of the DNA-recognition site for anti-DNA antibody inducing DNA-binding protein nucleobindin

抗 DNA 抗体诱导 DNA 结合蛋白核结合蛋白的 DNA 识别位点的探索

• 批准号: 07670528
• 负责人: KUBOTA Tetsuo
• 金额: $ 1.09万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670528/
• 关键词: nucleobindin; anti-DNA antibody; DNA-binding protein; autoimmunity; DNA結合性蛋白質; ヌクレオバインディン

We have achieved the following progresses in the studies on physiological activity and fine characteristics of nucleobindin.1. Administration of recombinant nucleobindin induced production of autoantibodies such as anti-DNA antibody even in normal BALB/c mice that have no genetical background of autoimmune disorders.2. Enhanced expression of nucleobindin mRNA was observed in the lymphoid organs of lupus-prone strains of mice such as MRL/lpr, C3H/gld, and male BXSB.3. Enhanced synthesis of nucleobindin in lupus-prone mice was demonstrated also in the protein level by immunohistochemical staining using an antibody produced against neclebindin peptide.4. A monoclonal antibody to human nucleobindin successfully immunoprecipitated native nucleobindin in the nuclear extract of human malignant lymphoma cell line Molt-4.5. The oligonucleotides which showed high binding affinity to human nucleobindin were selected from synthesized oligonucleotides containig 26-base-long random sequences. Some motif were repeatedly found in the selected oligonucleotides. Furthermore, such motif as CCACCACAATA and GTGTGGA were found in the up-stream region of IL-2 gene. It remains elusive how nucleobindin is involved in regulation of gene transcription.

在核结合素的生理活性和优良特性研究方面取得了以下进展.重组核结合蛋白的给药即使在没有自身免疫性疾病遗传背景的正常BALB/c小鼠中也能诱导自身抗体如抗DNA抗体的产生.在狼疮易感小鼠品系如MRL/lpr、C3H/gld和雄性BXSB的淋巴器官中观察到核结合蛋白mRNA的表达增强。在狼疮易感小鼠中，通过使用针对neclebindin肽产生的抗体的免疫组织化学染色，也在蛋白质水平上证明了核结合蛋白的合成增强。抗人核结合蛋白的单克隆抗体成功地免疫沉淀了人恶性淋巴瘤细胞系Molt-4.5的核提取物中的天然核结合蛋白。从人工合成的含有26个碱基的随机序列的寡核苷酸中筛选出与人核结合蛋白具有高亲和力的寡核苷酸。在所选的寡核苷酸中重复发现了一些基序。在IL-2基因的上游区域还发现了CCACCACAATA和GTGTGGA等基序。核结合蛋白如何参与基因转录的调控仍然是一个谜。

项目成果

Iizuka,H.et al.: "Enhanced expression of nucleobindin in lymphatic organs of lupus-prone mice" Lupus. (印刷中).

Iizuka, H. 等人：“狼疮易发小鼠淋巴器官中核结合蛋白的表达增强”狼疮（正在出版）。

Kanai,Y.et al.: "Induction of autoantibodies in normal mice by injection of nucleobindin and natural occurrence of antibodies in autoimmune MRL/lpr/lpr mice" Immunology Letters. 45. 35-42 (1995)

Kanai,Y.等人：“通过注射核结合素在正常小鼠中诱导自身抗体以及在自身免疫 MRL/lpr/lpr 小鼠中自然产生抗体”《免疫学快报》。

Kanai Y,Takeda O,Miura K,Amagai M,Kaneko T,Kubota T,Kanai Y,Tanuma S,Kurosawa Y: "Induction of autoantibodies in normal mice by injection of nucleobindin and natural occurrence of antibodies against nucleobindin in autoimmune MRL/lpr/lpr mice." Immunol Le

Kanai Y、Takeda O、Miura K、Amagai M、Kaneko T、Kubota T、Kanai Y、Tanuma S、Kurosawa Y：“通过注射核结合素在正常小鼠中诱导自身抗体，并在自身免疫 MRL/lpr 中自然产生针对核结合素的抗体

Kubota,T.: "Enhancement of oxidative cleavage of DNA by the binding sites of two anti-double stranded DNA antibodies." J.Boil.Chem.(in press).

Kubota,T.：“通过两种抗双链 DNA 抗体的结合位点增强 DNA 的氧化裂解。”