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Reticulon and CLIMP-63 control nanodomain organization of peripheral ER tubules

Guang Gao, Chengjia Zhu, Emma Liu, Ivan R Nabi
doi: https://doi.org/10.1101/550715
Guang Gao
Life Sciences Institute, University of British Columbia;
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  • For correspondence: gaoguang@mail.ubc.ca
Chengjia Zhu
Life Sciences Institute, University of British Columbia;
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  • For correspondence: c.zhu@alumni.ubc.ca
Emma Liu
Life Sciences Institute, University of British Columbia;
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  • For correspondence: e.liu@alumni.ubc.ca
Ivan R Nabi
University of British Columbia
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  • For correspondence: irnabi@mail.ubc.ca
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Abstract

The endoplasmic reticulum (ER) is an expansive, membrane-enclosed organelle composed of smooth peripheral tubules and rough, ribosome-studded central ER sheets whose morphology is determined, in part, by the ER-shaping proteins, reticulon and CLIMP-63, respectively. Here, STimulated Emission Depletion (STED) super-resolution microscopy shows that reticulon and CLIMP-63 also control the organization and dynamics of peripheral ER tubule nanodomains. STED imaging shows that lumenal ERmoxGFP, membrane Sec61βGFP, knock-in calreticulin-GFP and antibody-labeled ER resident proteins calnexin and derlin-1 are all localized to periodic puncta along the length of peripheral ER tubules that are not readily observable by diffraction limited confocal microscopy. Reticulon segregates away from and restricts lumenal blob length while CLIMP-63 associates with and increases lumenal blob length. Reticulon and CLIMP-63 also regulate the nanodomain distribution of ER resident proteins, being required for the preferential segregation of calnexin and derlin-1 puncta away from lumenal ERmoxGFP blobs. High-speed (40 ms/frame) live cell STED imaging shows that reticulon and CLIMP-63 control nanoscale compartmentalization of lumenal flow in peripheral ER tubules. Reticulon enhances and CLIMP-63 disrupts the local accumulation of lumenal ERmoxGFP at spatially defined sites along ER tubules. The ER shaping proteins reticulon and CLIMP-63 therefore control lumenal ER nanodomain dynamics, heterogeneity and interaction with ER resident proteins in peripheral ER tubules.

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Posted February 15, 2019.
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Reticulon and CLIMP-63 control nanodomain organization of peripheral ER tubules
Guang Gao, Chengjia Zhu, Emma Liu, Ivan R Nabi
bioRxiv 550715; doi: https://doi.org/10.1101/550715
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Reticulon and CLIMP-63 control nanodomain organization of peripheral ER tubules
Guang Gao, Chengjia Zhu, Emma Liu, Ivan R Nabi
bioRxiv 550715; doi: https://doi.org/10.1101/550715

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