Elucidation of the molecular mechanism of action of the yeast strongest DIT1 terminator and its application

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K07378
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Toyota Central R&D Lab., Inc.
• Principal Investigator: Matsuyama Takashi 株式会社豊田中央研究所, 社会システム研究領域 健康創出プログラム, 主任研究員 (90394882)
• Co-Investigator(Kenkyū-buntansha): 井沢 真吾 京都工芸繊維大学, 応用生物学系, 准教授 (10273517) ; 北川 孝雄 山口大学, 大学院医学系研究科, 助教(特命) (20614928)
• Co-Investigator(Renkei-kenkyūsha): IRIE Kenji 筑波大学, 医学医療系, 教授 (90232628)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 転写後発現制御 / ターミネーター / 出芽酵母

Outline of Final Research Achievements

In budding yeast, we analyzed the activation mechanism of the DIT1 terminator (DIT1t) with the highest activity to increase protein production of its upstream ORF. The cis sequences GUUCG/U were identified. Due to accumulation of deletion mutations that enhance the activity of DIT1t activation, an artificial DIT1t with about 1.5 times the activity of the wild type DIT1t was constructed.
Terminator activity of six genes involved in cell wall synthesis with GUUCG/U in the 3'-UTR was increased by overexpression of NAB6 and PAP1 genes. On the other hand, the nab6-disrupted strain was resistant to a cell wall synthetic inhibitor, caspofungin, and was sensitive to the drug congo red, which binds to another cell wall component chitin and causes growth inhibition. Taken together, it was suggested that a global regulation system using Nab6p-Pap1p complex as a hub might control cell wall synthesis.

Free Research Field

応用微生物学