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A novel plant actin-microtubule bridging complex regulates cytoskeletal and ER structure at ER-PM contact sites

Zang, Jingze; Klemm, Sandra; Pain, Charlotte; Duckney, Patrick; Bao, Zhiru; Stamm, Gina; Kriechbaumer, Verena; Bürstenbinder, Katharina; Hussey, Patrick J.; Wang, Pengwei

A novel plant actin-microtubule bridging complex regulates cytoskeletal and ER structure at ER-PM contact sites Thumbnail


Authors

Jingze Zang

Sandra Klemm

Charlotte Pain

Zhiru Bao

Gina Stamm

Verena Kriechbaumer

Katharina Bürstenbinder

Pengwei Wang



Abstract

In plants, the cortical endoplasmic reticulum (ER) network is connected to the plasma membrane (PM) through the ER-PM contact sites (EPCSs), whose structures are maintained by EPCS resident proteins and the cytoskeleton.1, 2, 3, 4, 5, 6, 7 Strong co-alignment between EPCSs and the cytoskeleton is observed in plants,1,8 but little is known of how the cytoskeleton is maintained and regulated at the EPCS. Here, we have used a yeast-two-hybrid screen and subsequent in vivo interaction studies in plants by fluorescence resonance energy transfer (FRET)-fluorescence lifetime imaging microscopy (FLIM) analysis to identify two microtubule binding proteins, KLCR1 (kinesin-light-chain-related protein 1) and IQD2 (IQ67-domain 2), that interact with the actin binding protein NET3C and form a component of plant EPCS that mediates the link between the actin and microtubule networks. The NET3C-KLCR1-IQD2 module, acting as an actin-microtubule bridging complex, has a direct influence on ER morphology and EPCS structure. Their loss-of-function mutants, net3a/NET3C RNAi, klcr1, or iqd2, exhibit defects in pavement cell morphology, which we suggest is linked to the disorganization of both actin filaments and microtubules. In conclusion, our results reveal a novel cytoskeletal-associated complex, which is essential for the maintenance and organization of cytoskeletal structure and ER morphology at the EPCS and for normal plant cell morphogenesis.

Citation

Zang, J., Klemm, S., Pain, C., Duckney, P., Bao, Z., Stamm, G., …Wang, P. (2021). A novel plant actin-microtubule bridging complex regulates cytoskeletal and ER structure at ER-PM contact sites. Current Biology, 31(6), 1251-1260.e4. https://doi.org/10.1016/j.cub.2020.12.009

Journal Article Type Article
Acceptance Date Dec 9, 2020
Online Publication Date Feb 15, 2021
Publication Date Mar 22, 2021
Deposit Date Feb 18, 2021
Publicly Available Date Feb 15, 2022
Journal Current Biology
Print ISSN 0960-9822
Publisher Cell Press
Peer Reviewed Peer Reviewed
Volume 31
Issue 6
Pages 1251-1260.e4
DOI https://doi.org/10.1016/j.cub.2020.12.009

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