https://scholars.lib.ntu.edu.tw/handle/123456789/159878
Title: | 生長休止基因7於細胞週期蛋白依賴型激酶5及其激動分子p35複合體之研究 The study of Growth arrest-specific 7 Involves in Cyclin-dependent kinase 5/p35 Kinase complex |
Authors: | 鍾岱璉 Chung, Tai-Lien |
Keywords: | 生長休止基因七;Gas7 | Issue Date: | 2005 | Abstract: | 細胞週期蛋白依賴型激酶5(Cdk5)及其激動分子p35主要調控老鼠腦部發育的過程。近來發現,Cdk5/p35複合體位於細胞高基氏體中,並且負責液泡運送的調控,但是該分子機制尚不清楚。生長休止基因七 (Gas7)被發現大量表現在細胞生長休止階段 (G0 phase),並已知主要表現在腦部。CBP1是Gas7羧基(COOH)端的交互作用分子,且其序列和p35的交互作用分子,稱為C48,有高度的相似性,但是目前對於該交互分子---C48,的生物功能仍不了解。此篇論文主要是探討: Gas7是否會藉由CBP1和Cdk5及p35形成複合體,並且扮演維持高基氏體的正常結構並進一步調控其細胞功能,如液泡運送的角色。利用谷胱甘肽S-轉移(GST)的沉澱分析(GST pull down assay),得知Gas7無法直接與Cdk5及p35交互作用,而是必須藉由CBP1。經由海馬回初代細胞的免疫染色發現,CBP1、Gas7、Cdk5和p35都有表現在細胞高基氏體。並且在老鼠腦部的發育時期,藉由蔗糖梯度法分離高基氏體(Golgi enriched sucrose gradients),Gas7和p35會主要表現在高基氏體並和Cdk5及actin形成複合物。進一步利用定點突變蛋白VSVG-ts045-GFP蛋白運送分析,在大量表現CBP1的細胞中,會造成液泡從內質網運送到高基氏的過程有延後的現象。總結來說,Gas7及CBP1會與Cdk5及p35形成複合體,並且同樣會位於細胞高基氏體中,而CBP1似乎參與細胞液泡運送調控的機制。 Cyclin-dependent kinase 5 (Cdk5), a serine/threonine kinase, which is dependent on association with its neuronal specific activator, p35, is required for proper development of a mouse brain. Recently, the Cdk5-p35 kinase has been detected in the Golgi apparatus and is a factor in the regulation of membrane traffic, although the action of Cdk5-p35 in the Golgi apparatus is largely unclear. The growth arrest-specific 7 (Gas7) is identified in G0 phase and expressed preferentially in the brain. Gas7 C’-terminal Binding Protein, CBP1, is highly homologous with p35-interacting protein, but its biological function is not understood. Moreover, Gas7 is associated with actin assembly that is responsible for protein transport in Golgi apparatus. This study will determine the complex formation of Gas7, CBP1, p35 and Cdk5 tetra-complex, and their roles in Golgi architecture and Golgi-mediated cellular functions including vesicle transport. In protein-protein relationships, no direct interaction been detected between Gas7 and p35/Cdk5 kinase without mediation by CBP1. Preliminary results indicate that the pattern of Golgi distribution by analyzing specific Golgi markers of sucrose gradient prepared from Gas7 wild-type developing mouse brains reveals Gas7 and p35 are predominantly expressed in Golgi-enriched fractions. Moreover, the tetra-complex formation of Gas7, actins and Cdk5/p35, exists in Golgi-enriched microsomal fractions purified from the developing mouse brain. In the VSVG-ts045-GFP transport assay, CBP1 delays the vesicle transportation from the ER to the Golgi apparatus. Collectively, these results indicate that CBP1, a key component for Gas7 and p35/CDK5 complex formation, is not only predominantly expressed in the Golgi complex but also plays a role in protein trafficking. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/51349 | Other Identifiers: | en-US |
Appears in Collections: | 分子醫學研究所 |
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ntu-94-R92448008-1.pdf | 23.31 kB | Adobe PDF | View/Open |
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