Isolation, characterization of represser protein in expression of human DLST gene.

人 DLST 基因表达中阻遏蛋白的分离、表征。

• 批准号: 07680694
• 负责人: MATUDA Sadayuki
• 金额: $ 1.41万
• 依托单位: National Institute of Fitness & Sports in Kanoya
• 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-07680694/
• 关键词: Dihydrolipoamide succinyltransferase; alpha-ketoglutarate dehydrogenase complex; regulation of gene expression; analysis of function of promoter region; プロモーター領域

In this study, we analyzed the function in expression of human dihydro lipoamide succinyltransferase gene. The enzyme is a core-component of 2-oxoglutarate dehydrogenase complex and plays a role in energy production in mitrochondria. We have isolated the genomic DNA for the DLST gene and determined the entire nucleotide sequence. Sequence analysis of the promoter-regulatory region of the DLST gene showed the presence of a CAAT-box-like sequence but the presence of a TATA-box-like sequence was not evidenced. Also located in this region were sequences resembling glucocorticoid-responsive and cAMP-responsive elements, and an Spl binding site. Thus, the DLST gene is a typical housekeeping gene. Gene group of this type, possessing multiple GC boxes but no TATA box, is activated by transcription factor Spl or Spl-related proteins. In this study, however, we found that the DLST gene is not activated by Spl or Spl-related proteins. A region-35 to-12 upstream from the transcription site is required for the transcription of the DLST gene. Furthermore, we found that the a region +796 to +964 in the second intron strongly supresses the expression of the DLST gene. The DLST gene may be regulated by the combination of the transcrption-activation region-35 to-12 and of the transcription-supression region +796 to +964.

本研究分析了人二氢硫辛酰胺琥珀酰转移酶基因的表达功能。该酶是2-酮戊二酸脱氢酶复合物的核心组分，在线粒体的能量产生中起作用。我们已经分离了DLST基因的基因组DNA，并确定了整个核苷酸序列。DLST基因的启动子调控区的序列分析表明存在CAAT盒样序列，但TATA盒样序列的存在没有得到证明。同样位于该区域的是类似糖皮质激素响应和cAMP响应元件的序列，以及Spl结合位点。因此，DLST基因是典型的管家基因。这类基因群具有多个GC盒，但没有TATA盒，由转录因子Spl或Sl相关蛋白激活。然而，在这项研究中，我们发现DLST基因不被Spl或Spl-related蛋白激活。转录位点上游的-35至-12区是DLST基因转录所必需的。此外，我们发现第二内含子中的a区域+796至+964强烈抑制DLST基因的表达。DLST基因可能受转录激活区-35至-12和转录抑制区+796至+964的组合调控。

项目成果

T.Takase, K.Nakano, S.Ohta, S.Nakagawa and S.Matuda: "Different distribution of dihydrolipoamide succinyltransferase, dihydrolipoamide acetyl-transferaseand ATP synthase beta-subunit in monkey brain." in vivo. Vol. 10. 495-502 (1996)

T.Takase、K.Nakano、S.Ohta、S.Nakakawa 和 S.Matuda：“二氢硫辛酰胺琥珀酰转移酶、二氢硫辛酰胺乙酰转移酶和 ATP 合成酶 β 亚基在猴脑中的不同分布。”

T.Takase et al.: "Different distribution of dihydrolipoamide succinyltransferase,dihydrolipoamide acetyltransferase and ATP synthase β-subunit in monkey brain." in vivo. 10. 495-502 (1996)

T. Takase 等人：“猴脑中二氢硫辛酰胺琥珀酰转移酶、二氢硫辛酰胺乙酰转移酶和 ATP 合酶 β 亚基的不同分布。”10. 495-502 (1996)。

T.Takase et al.: "Different distribution of dihydrolipoamide succinyltransferase,dihydrolipoamide acetyltransferase and ATP synthase β-subuntit in monkey brain." in vivo. 10. 495-502 (1996)

T. Takase 等人：“猴脑中二氢硫辛酰胺琥珀酰转移酶、二氢硫辛酰胺乙酰转移酶和 ATP 合酶 β 亚基的不同分布。”10. 495-502 (1996)。

S.Matuda et al.: "A dihydrolipoamide succinyltransferase related protein is present on the plasma membrane in skeletal muscle." FEBS Letter. (印刷中). (1996)

S. Matuda 等人：“骨骼肌质膜上存在二氢硫辛酰胺琥珀酰转移酶相关蛋白”（FEBS Letter）（1996 年）。

T.Takase et al.: "Differential distribution of dihydrolipoamide succinyltransferase,dihydrolipoamide acetyltransferase and ATP synthase β-subunit in monkey brain." in vivo. (印刷中). (1996)

T. Takase 等人：“猴脑中二氢硫辛酰胺琥珀酰转移酶、二氢硫辛酰胺乙酰转移酶和 ATP 合酶 β 亚基的差异分布”（正在出版）。