Dissection of nuclear envelope assembly by constitutive approach using artificial beads in living cells

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K06238
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 44010:Cell biology-related
• Research Institution: National Institute of Information and Communications Technology
• Principal Investigator: Kobayashi Shouhei 国立研究開発法人情報通信研究機構, 未来ICT研究所フロンティア創造総合研究室, 研究マネージャー (40425765)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 核膜 / 核膜形成 / 人工ビーズ / 核膜孔複合体 / importin-beta / BAF / Live-CLEM

Outline of Final Research Achievements

This research aims to elucidate the mechanism of nuclear envelope formation in human somatic cells. To this end, we developed an experimental system in which nuclear membrane-like structures with nuclear pore complexes (NPC) can be assembled around a biomolecule-coated bead in a living cell. Immunofluorescence staining and live correlative light-electron microscopy (Live-CLEM) revealed that the beads coated with effector molecules, such as importin-beta, efficiently assembled NE-like structures with several components of NPC. In contrast, the beads coated with barrier-to-autointegration factor (BAF), a factor to assemble the LEM domain NE proteins, also efficiently assembled the NE-like structures containing emerin, but not the NPC components. These results suggest that assembly of the NPC is a mutually independent process with that of the LEM domain proteins.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

本研究では、従来までカエル卵抽出液を用いた方法でしか実現困難であった核膜孔を有する核膜構造の人工形成を、生きたままのヒト培養細胞の中で実現した。このことは、より生細胞内環境に近い条件下での核膜形成機構の解析を可能とするため、細胞核研究分野の進展に大きく貢献すると思われる。また、本研究成果は、核膜形成以外の細胞内現象の構成的アプローチでの解析技術としての発展も期待されるため、多くの研究分野に大きなインパクトを与え得るものと考えている。