Exploration of Glucosylceramide Transport Factors Using CRISPR Library

2021 Fiscal Year Final Research Report

• Project/Area Number: 17K07357
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: Yamaji TOshiyuki 国立感染症研究所, 細胞化学部, 室長 (50332309)
• Project Period (FY): 2017-04-01 – 2022-03-31
• Keywords: スフィンゴ糖脂質 / ゲノム編集 / スクリーニング / ゴルジ体

Outline of Final Research Achievements

In this study, we used CRISPR knockout library to search for novel factors affecting glycolipid transport. We succeeded in identifying genes involved in glycolipid metabolisms with high coverage. We succeeded in identifying SYS1, a multi-spanning transmembrane protein localized in the Golgi apparatus, in Vero cells. Knockout of SYS1 gene reduced Gb3 and instead remarkably increased GlcCer. The SYS1 knockout cells also showed a change in the morphology of the Golgi apparatus from an elongated laminar to an aggregated form, and dot-like structures of a trans-Golgi network marker TGN46 around the Golgi apparatus. This indicates that SYS1 regulates glycan biosynthesis through its involvement in the Golgi apparatus and trans-Golgi network trafficking.

Free Research Field

糖鎖生物学　脂質生物学　感染症学

Academic Significance and Societal Importance of the Research Achievements

スフィンゴ糖脂質は、高次機能や疾患に深く関与することが知られており、生体に欠かせない脂質である。スフィンゴ糖脂質の生合成酵素はほぼ同定されているものの、細胞内輸送因子に関しては未解明な点が残されており、代謝・輸送に関する因子の同定は重要な課題である。本研究により、ゴルジ体における糖鎖生合成の制御に関して重要な知見を得ており、糖鎖生物学及び細胞生物学的分野における学術的意義を示すものである。