Structural determinants of REMORIN nanodomain formation in anionic membranes

Legrand, Anthony; G-Cava, Daniel; Jolivet, Marie-Dominique; Décossas, Marion; Lambert, Olivier; Bayle, Vincent; Jaillais, Yvon; Loquet, Antoine; Boudsocq, Marie; Habenstein, Birgit; Tirado, Marisela Vélez; Mongrand, Sébastien

doi:10.1016/j.bpj.2022.12.035

Cited by 9 publications

(23 citation statements)

References 57 publications

(102 reference statements)

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“…The Arabidopsis thaliana calcium-dependent protein kinase 3 (AtCPK3) was previously shown to phosphorylate group 1 REMORINs (Mehlmer et al, 2010; Perraki et al, 2018) and to restrict PVX cell-to-cell movement in a REMORIN-dependent manner (Perraki et al, 2018). Liquid-state NMR spectroscopy showed that the addition of AtCPK3, which phosphorylates StREM1.3 N-terminal domain in vitro (Legrand et al, 2022; Perraki et al, 2018) (Figure S2), induced chemical shift perturbations on several residues without altering the intrinsically disordered nature of StREM1.3 1-116 (Figures 1B and 1C). The chemical shifts of the amide groups are very sensitive to the local chemical environment of the concerned residues.…”

Section: Resultsmentioning

confidence: 99%

“…StREM1.3 1-116 was obtained as previously described (Legrand et al, 2022). Shortly, StREM1.3 1-116 was expressed in BL21-DE3 cells in minimal medium with 13 C-labelled glucose and 15 NH 4 Cl, by addition of 1 mM IPTG at OD 600 = 0.6-0.8 and incubation at 37°C for 3h.…”

Section: Methodsmentioning

confidence: 99%

“…Finally, StREM1.3 1-116 was desalted again against 10 mM HEPES, 50 mM NaCl 0.02%, NaN 3 pH=7.5. AtCPK3-GST recombinant protein was produced in BL21-DE3-pLys and purified as previously reported (Legrand, et al, 2022). As a negative control, AtCPK3-GST was inactivated by heating at 95°C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…In vitro phosphorylation of 0.5 mM of StREM1.3 1-116 by AtCPK3 was done as previously published (Legrand et al, 2022). For NMR analysis, the StREM1.3 1-116 sample was adjusted to 10 mM MgCl 2 , 1 mM CaCl 2 , 1 mM DTT and 3 mM ATP.…”

Section: Methodsmentioning

confidence: 99%

“…Group 1 REMORINs NDs are proposed to be liquid-ordered and enriched in sterols and saturated lipids (Demir et al, 2013; Gronnier et al, 2017; Mamode Cassim et al, 2019; Raffaele et al, 2009). Their native organization is dictated by direct interactions with anionic phospholipids (Gronnier et al, 2017; Legrand et al, 2019; Legrand et al, 2022; Perraki et al, 2012), and can be modulated by post-translational modifications such as S-acylation (Fu et al, 2018; Konrad et fal., 2014) and phosphorylation (Perraki et al, 2018). Group 1 REMORINs regulate cell-to-cell movement of viruses belonging to diverse genus (Cheng et al, 2020; Fu et al, 2018; Huang et al, 2019; Raffaele et al, 2009; Rocher et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

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The actin depolymerizing factor StADF2 alters StREM1.3 plasma membrane nanodomains to inhibit thePotato Virus X

Jolivet¹,

Gouguet²,

Legrand³

et al. 2023

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The dynamic regulation of the plasma membrane (PM) organization at the nanoscale emerged as a key element shaping the outcome of host-microbe interactions. Protein organization into nanodomains (ND) is often assumed to be linked to the activation of cellular processes. In contrast, we have previously shown that the phosphorylation of the Solanum tuberosum REM1.3 (StREM1.3) N-terminal domain disperses its native ND organization and promotes its inhibitory effect on Potato Virus X (PVX) cell-to-cell movement. Here, we show that the phosphorylation of StREM1.3 modify the chemical environment of numerous residues in its intrinsically-disordered N-terminal domain. We leveraged exploratory screens to identify potential phosphorylation-dependent interactors of StREM1.3. Herewith, we uncovered uncharacterized regulators of PVX cell-to-cell movement, linking StREM1.3 to autophagy, water channels and the actin cytoskeleton. We show that the Solanum tuberosum actin depolymerizing factors 2 (StADF2) alters StREM1.3 NDs and limits PVX cell-to-cell movement in a REMORIN-dependent manner. Mutating a conserved single residue reported to affect ADFs affinity to actin inhibits StADF2 effect on StREM1.3 ND organization and PVX cell-to-cell movement. These observations provide functional links between the organization of plant PM and the actin cytoskeleton and suggests that the alteration of StREM1.3 ND organization promotes plant anti-viral responses. We envision that analogous PM re-organization applies for additional signaling pathways in plants and in other organisms.

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