SIGNAL TRANSDUCTION AND CYTOSKELTON

信号转导和细胞骨架

• 批准号: 08660410
• 负责人: KAWAMUKAI Makoto
• 金额: $ 1.34万
• 依托单位: SHIMANE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660410/
• 关键词: RAS; cAMP; Signal transduction; fission yeast; CAP; cytoskelton; RAS; cAMP

The adenylyl cyclase - associated protein, CAP,was first identified in Saccharomyces cerevisiae, and succeedingly, its homologs were found in many eukaryotes ranging from yeasts to humans. Yeast CAPs have been found to have three functions : the N - terminal domain as the region for binding adenylyl cyclase, the C - terminal domain as an actin binding domain, and the central proline - rich domainas the SH3 (Src Homology 3) binding region. Here, we report the identification of a cDNA and genomic DNA encoding a CAP homolog from mushroom Lentinus edodes. Southern blotting results suggest it to be a single - copy gene. The L.edodes cap gene contains 8 introns and an open reading frame encoding a 518 amino acid protein. The C - terminal domain shares higher homology (39 - 46 % identity) compared with the N - terminal domain (27 - 35 % identity) with yeast CAPs. Expression of the L.edodes CAP in both S.pombe and in S.cerevisiae complemented defects associated with loss of the C - terminal domain function of endogenous CAP.By using yeast two - hybrid assay, we demonstrated the interaction between the L.edodes CAP and S.pombe actin. This result andfunctional complementation test indicate CAP from mushroom is conserved in terms of the C - terminal function of CAP.

腺苷酸环化酶相关蛋白 CAP 首次在酿酒酵母中被发现，随后在从酵母到人类的许多真核生物中发现了其同源物。酵母CAP被发现具有三种功能：N端结构域作为腺苷酸环化酶的结合区域，C端结构域作为肌动蛋白结合结构域，以及中央富含脯氨酸的结构域作为SH3（Src Homology 3）结合区域。在这里，我们报告了从蘑菇香菇中编码 CAP 同源物的 cDNA 和基因组 DNA 的鉴定。 Southern印迹结果表明它是一个单拷贝基因。 L.edodes cap基因含有8个内含子和一个编码518个氨基酸的蛋白质的开放阅读框。与酵母CAP的N端结构域（27-35%同一性）相比，C端结构域具有更高的同源性（39-46%同一性）。 L.edodes CAP在S.pombe和S.cerevisiae中的表达补充了与内源CAP的C末端结构域功能丧失相关的缺陷。通过使用酵母双杂交实验，我们证明了L.edodes CAP和S.pombe肌动蛋白之间的相互作用。该结果和功能互补测试表明蘑菇的CAP在CAP的C端功能方面是保守的。

项目成果

川向 誠: "アデニル酸シクラーゼ結合タンパク質(AP)とそのホモログ" 王化学. 68. 31-35 (1996)

Makoto Kawamukai：“腺苷酸环化酶结合蛋白（AP）及其同源物”Oh Kagaku。68. 31-35（1996）

Suzuki Kengoら: "Analysis of the decapreyl diphoephate synthese(olps)gone in fission yeast suggeots a rok of uhigui mine on an infictialant" J.Biochem. 121. 496-505 (1997)

Suzuki Kengo 等人：“对裂殖酵母中的十癸酰二磷酸合成酶 (olps) 的分析表明，在感染者中存在 Uhigui 矿” J.Biochem. 121. 496-505 (1997)

Kazunori Okada他: "Polypronyl diphosphitesynthase essontially defines the leryth of the slde chain of ubiqunone" Biochimica et Biophysica Acta. 1302. 217-223 (1996)

Kazunori Okada 等人：“聚丙酰二磷酸合成酶本质上定义了泛醌的链的结构”Biochimica et Biophysicala Acta. 1302. 217-223 (1996)