SUMOylation is involved in the regulation of cell cycle check point

2016 Fiscal Year Final Research Report

• Project/Area Number: 25440134
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Plant molecular biology/Plant physiology
• Research Institution: Kumamoto University
• Principal Investigator: Ishida Takashi 熊本大学, 国際先端科学技術研究機構, 助教 (00462656)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: チェックポイントタンパク質 / シロイヌナズナ / 細胞周期 / SUMO

Outline of Final Research Achievements

In this study, we analyzed cell cycle regulation of plant cell, especially focusing on the chromosome check point proteins. First, we generated knock-down lines with artificial microRNA technology, and observed delayed growth and abnormal morphology phenotypes. In addition, we also generated knock-out mutants with the CRISPR / Cas9 systems. As expected, we could not obtained homozygote seedlings, suggesting that the loss-of-function mutants are embryo lethal. Next, we substitute the amiRNA target sequence of the checkpoint protein encoding gene to make it insensitive to amiRNA, then transformed into the knock-down line. The introduction of the fragment could rescue the abnormalities of mutants, indicating that the fragment was functional. In contrast, an additional SUMO-null mutation spoiled the functionalities of the genome fragment, suggesting that the SUMOylation is required for the function. We also examined the protein-protein interactions with the SUMO E3 ligase by co-IP assays.

Free Research Field

植物細胞生物学