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First published online 15 July 2008
doi: 10.1242/jcs.033597

Journal of Cell Science 121, 2540-2554 (2008)
Published by The Company of Biologists 2008
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Research Article

Live cell imaging and electron microscopy reveal dynamic processes of BAF-directed nuclear envelope assembly

Tokuko Haraguchi1,2,*, Tomoko Kojidani1, Takako Koujin1, Takeshi Shimi1, Hiroko Osakada1, Chie Mori1, Akitsugu Yamamoto3 and Yasushi Hiraoka1,2,4

1 CREST Research Project, Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan
2 Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka 560-0043, Japan
3 Nagahama Institute of Bio-Science and Technology, 1266 Tamura-cho, Nagahama 526-0829, Japan
4 Graduate School of Frontier Biosciences, Osaka University 1-3 Suita 565-0871, Japan

* Author for correspondence (e-mail: tokuko{at}nict.go.jp)

Accepted 19 May 2008

Assembly of the nuclear envelope (NE) in telophase is essential for higher eukaryotic cells to re-establish a functional nucleus. Time-lapse, FRAP and FRET analyses in human cells showed that barrier-to-autointegration factor (BAF), a DNA-binding protein, assembled first at the distinct `core' region of the telophase chromosome and formed an immobile complex by directly binding with other core-localizing NE proteins, such as lamin A and emerin. Correlative light and electron microscopy after live cell imaging, further showed that BAF formed an electron-dense structure on the chromosome surface of the core, close to spindle microtubules (MTs) prior to the attachment of precursor NE membranes, suggesting that MTs may mediate core assembly of BAF. Disruption of the spindle MTs consistently abolished BAF accumulation at the core. In addition, RNAi of BAF eliminated the core assembly of lamin A and emerin, caused abnormal cytoplasmic accumulation of precursor nuclear membranes and resulted in a significant delay of NE assembly. These results suggest that the MT-mediated BAF accumulation at the core facilitates NE assembly at the end of mitosis.

Key words: Barrier-to-autointegration factor, Chromatin, Emerin, Lamin A, Nuclear envelope, Microtubule


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JCS 2008 121: 1501. [Full Text]  





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