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bioRxiv · 10.1101/2025.05.20.655051

Chloroplast cpHsc70-1 interacts with VIPP1 C-terminal tail and controls VIPP1 oligomer disassembly in thylakoid membrane remodeling

Li, D.·Gachie, S. W.·Ozawa, S.-I.·Scholz, M.·Hippler, M.·Sakamoto, W.
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Abstract

Oxygenic photosynthetic organisms depend on the thylakoid membranes (TMs) for light-driven energy conversion. Recent studies on TM homeostasis (thylakostasis) have highlighted the essential role of the TM remodeling protein VIPP1 (Vesicle Inducing Protein in Plastids 1). As a member of the ESCRT-III/PspA/VIPP1 superfamily, VIPP1 forms large ring- and filament-like homo-oligomeric structures that exhibit a membrane remodeling activity. The oligomerization status was proposed to be modulated by the intrinsically disordered C-terminal tail (Vc), whereas its functional role remained unclear. Notably, this Vc region is conserved not only in photosynthetic VIPP1 but also in the PspA proteins of extremophilic species, implicating its role in membrane stress responses. To investigate the role of the Vc region in VIPP1 assembly, we performed co-immunoprecipitation assays in Arabidopsis chloroplasts and identified chloroplast-localized HSP70 proteins as major interactors. Among the two isoforms, cpHsc70-1 was found to be specifically required for modulating VIPP1 oligomeric assembly and dynamics in response to heat stress. Genetic analyses revealed that cpHsc70-1 facilitates the disassembly of VIPP1 oligomers, similarly to Vps4 ATPase in ESCRT-III; loss of either the Vc region or cpHsc70-1 impaired VIPP1 disassembly, resulting in more static oligomeric structures. Furthermore, cpHsc70-1 exhibited a broader role in chloroplast proteostasis, as the cphsc70-1 mutant showed impaired accumulation of GFP-fusion proteins. Together, our findings uncover a crucial crosstalk between proteostasis and thylakostasis in chloroplasts, coordinated by cpHsc70-1 and VIPP1 in response to membrane stress. Significance StatementOrganisms performing oxygen-evolving photosynthesis rely on the thylakoid membranes (TMs) for light-driven energy conversion. We reveal that VIPP1, a membrane remodeling protein forming dynamic oligomeric structures, is crucial for TM maintenance. Its intrinsically disordered C-terminal region (Vc) and the heat shock protein cpHsc70-1 coordinate VIPP1 disassembly during stress in Arabidopsis chloroplasts. Loss of either component causes defects in membrane remodeling and proteostasis. Our findings uncover a fundamental mechanism connecting membrane integrity and protein quality control, ensuring chloroplast resilience against environmental stress.

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Li, D., Gachie, S. W., Ozawa, S.-I., Scholz, M., Hippler, M., Sakamoto, W.. 2025-05-23. Chloroplast cpHsc70-1 interacts with VIPP1 C-terminal tail and controls VIPP1 oligomer disassembly in thylakoid membrane remodeling. https://doi.org/10.1101/2025.05.20.655051

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