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Genes to Cells (2005) 10, 989-999. doi:10.1111/j.1365-2443.2005.00894.x
© 2005 Blackwell Publishing or its licensors

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NSF/SNAPs and p97/p47/VCIP135 are sequentially required for cell cycle-dependent reformation of the ER network

Fumi Kano1, Hisao Kondo2,3,4, Akitsugu Yamamoto5, Yayoi Kaneko3, Keiji Uchiyama2,3, Nobuko Hosokawa6, Kazuhiro Nagata6 and Masayuki Murata1,*

1 Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Tokyo 153-8902, Japan
2 Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 2XY, UK s
3 Mitsubishi Institute of Life Science, Machida, Tokyo 194-8511, Japan
4 PRESTO, Japan Science and Technology Corporation, Japan
5 Cellular Structure Laboratory, Cell Biology Program, Nagahama Institute of Bioscience and Technology, Shiga 526-0829, Japan
6 Department of Molecular and Cellular Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan

The endoplasmic reticulum (ER) has a characteristic polygonal structure with hallmark three-way junctions. In a previous paper, we reconstituted the disruption of the pre-existing ER network using mitotic cytosol from HeLa cells in streptolysin O (SLO)-permeabilized CHO-HSP cells (stably expressing GFP-HSP47). In addition, we found that interphase cytosol induced reformation of the disrupted ER network into a continuous network structure. Here, we show that the reformation of the ER network is accomplished through two sequential fusion reactions. The first process is mediated by NSF/{alpha} and {gamma}-SNAPs, and involves the generation of typical membranous intermediate structures that connect the disrupted ER tubules. A subsequent fusion is mediated by p97/p47/VCIP135, which has been shown to be required for homotypic fusion events in Golgi cisternae regrowth after mitosis. In addition, we also found that both fusion processes involve the t-SNARE, syntaxin 18.


Communicated by: Yoshinori Ohsumi

* Correspondence: E-mail: mmurata{at}bio.c.u-tokyo.ac.jp




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