In vitro assembly of nuclear envelope in tobacco cultured cells.

Abstract:

:The coordinated regulation of the nucelar envelope (NE) reassembly during cell division is an essential event. However, there is little information on the molecular components involved in NE assembly in plant cells. Here we developed an in vitro assay of NE assembly using tobacco BY-2 cultured cells. To start the NE assembly reaction, the demembranated nuclei and the S12 fraction (cytosol and microsomes) were mixed in the presence of GTP and ATP nucleotides. Time-course analysis indicated that tubule structures were extended from the microsomal vesicles that accumulated on the demembranated nuclei, and finally sealed the NE. Immunofluorescence confirmed that the assembled membrane contains a component of nuclear pore complex. The efficiency of the NE assembly is significantly inhibited by GTPγS that suppresses membrane fusion. This in-vitro assay system may elucidate the role of specific proteins and provide important insights into the molecular machinery of NE assembly in plant cells.

journal_name

Nucleus

journal_title

Nucleus (Austin, Tex.)

authors

Tamura K,Ueda H,Hara-Nishimura I

doi

10.1080/19491034.2021.1930681

keywords:

["BY-2","confocal microscopy","endoplasmic reticulum","nuclear envelope","nucleus"]

subject

Has Abstract

pub_date

2021-12-01 00:00:00

pages

82-89

issue

1

eissn

1949-1034

issn

1949-1042

journal_volume

12

pub_type

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