Cell biological analysis of membrane protein induced by glucocorticoid

2014 Fiscal Year Final Research Report

• Project/Area Number: 24592808
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Nagasaki University
• Principal Investigator: OKAMOTO Kuniaki 長崎大学, 医歯薬学総合研究科(歯学系), 准教授 (10311846)
• Co-Investigator(Kenkyū-buntansha): TSUKUBA Takayuki 長崎大学, 医歯薬学総合研究科(歯学系), 教授 (30264055) ; NISHISHITA Kazuhisa 長崎大学, 医歯薬学総合研究科(歯学系), 助教 (20237697) ; SAKAI Eiko 長崎大学, 医歯薬学総合研究科(歯学系), 助教 (10176612)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: ステロイド / エンドソーム・リソソーム / オートファジー / 骨芽細胞

Outline of Final Research Achievements

Transcription Factor EB (TFEB) is a master gene that activates the formation of autophagozomes, and upregulates the expression of lots of lysosomal enzymes. Among them, dexamethasone-induced transcript (DEXI) is also one of proteins that the expression increases. It is reported that DEXI is a glucocorticoid-induced gene upregulated in Emphysema. However, it is not unknown about its localization or functions. Therefore, to examine the localization of DEXI, we amplified full length gene of DEXI from mouse cDNA by using polymerase chain reaction and constructed DEXI expression vector fused with GFP. Although transfected this construct in MC3T3-E1 and HeLa cells, no or only few expression of DEXI was observed by western blot. The reason why DEXI did not express may be that the protein is very small compared with GFP protein. Then, we changed GFP into FLAG tag. At the same time, we also amplified TFEB gene from mouse and constructed TFEB expression vector fused with FLAG tag.

Free Research Field

歯科薬理学