Structural and Functional Insights into the Cryoprotection of Membranes by the Intrinsically Disordered Dehydrins

Clarke, Matthew W.; Boddington, Kelly F.; Warnica, Josephine M.; Atkinson, John; McKenna, Sarah E; Madge, Jeffrey; Barker, Christine H.; Graether, Steffen P.

doi:10.1074/jbc.m115.678219

Cited by 47 publications

(76 citation statements)

References 69 publications

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“…This reversible binding corresponds well to that observed for several other intrinsically disordered proteins upon binding specific protein targets (Jao et al, 2004;. Moreover, a seemingly analogous helix induction was observed recently by NMR for the dehydrin K 2 upon binding to SDS micelles (Clarke et al, 2015), pointing to the possibility that this mode of membrane interaction is a general feature of the conserved K segments.…”

Section: Nmr-monitored Folding Of the Conserved K Segment Flanked By supporting

confidence: 87%

“…The consequence of this membrane interaction is a decrease of the main lipid phase transition by 2.5°C in vitro (Eriksson et al, 2011), intriguingly matching the decreased survival temperature of 3°C observed upon Lti30 overexpression in Arabidopsis (Puhakainen et al, 2004). Consistent effects on the membrane phase transition are reported for the dehydrin K 2 (Clarke et al, 2015). To achieve membrane binding, Lti30 comprises not less than six copies of the archetypical K segment (Fig.…”

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confidence: 66%

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Membrane-induced folding of the plant-stress protein Lti30.

et al. 2016

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Dehydrins are disordered proteins that are expressed in plants as a response to embryogenesis and water-related stress. The molecular function and structural action of the dehydrins are yet elusive, but increasing evidence points to a role in protecting the structure and functional dynamics of cell membranes. An intriguing example is the cold-induced dehydrin Lti30 that binds to membranes by its conserved K segments. Moreover, this binding can be regulated by pH and phosphorylation and shifts the membrane phase transition to lower temperatures, consistent with the protein's postulated function in cold stress. In this study, we reveal how the Lti30-membrane interplay works structurally at atomic level resolution in Arabidopsis (Arabidopsis thaliana). Nuclear magnetic resonance analysis suggests that negatively charged lipid head groups electrostatically capture the protein's disordered K segments, which locally fold up into a-helical segments on the membrane surface. Thus, Lti30 conforms to the general theme of structure-function relationships by folding upon binding, in spite of its disordered, atypically hydrophilic and repetitive sequence signatures. Moreover, the fixed and well-defined structure of the membrane-bound K segments suggests that dehydrins have the molecular prerequisites for higher level binding specificity and regulation, raising new questions about the complexity of their biological function.

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Section: Nmr-monitored Folding Of the Conserved K Segment Flanked By supporting

confidence: 87%

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confidence: 66%

Membrane-induced folding of the plant-stress protein Lti30.

et al. 2016

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“…Experimental evidence for a stabilizing effect on membranes under stress conditions has so far been obtained for two mitochondrial and two plastidic LEA_4 (PF02987) proteins as well as one cytosolic LEA_5 (PF00477) protein and one dehydrin. The mitochondrial proteins are LEAM from pea ( Pisum sativum ) seeds and AfrLEA3m from the brine shrimp Artemia franciscana , the plastidic proteins are COR15A and COR15B from Arabidopsis , the cytosolic LEA_5 protein is AfrLEA2 from A. franciscana , and the dehydrin is K 2 from Vitis riparia . All six proteins are disordered under fully hydrated conditions and most fold at least partially into α‐helices when completely dried .…”

Section: Introductionmentioning

confidence: 99%

Folding of intrinsically disordered plant LEA proteins is driven by glycerol‐induced crowding and the presence of membranes

et al. 2017

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Late embryogenesis abundant (LEA) proteins are related to cellular dehydration tolerance. Most LEA proteins are predicted to have no stable secondary structure in solution, i.e., to be intrinsically disordered proteins (IDPs), but they may acquire α-helical structure upon drying. In the model plant Arabidopsis thaliana, the LEA proteins COR15A and COR15B are highly induced upon cold treatment and are necessary for the plants to attain full freezing tolerance. Freezing leads to increased intracellular crowding due to dehydration by extracellular ice crystals. In vitro, crowding by high glycerol concentrations induced partial folding of COR15 proteins. Here, we have extended these investigations to two related proteins, LEA11 and LEA25. LEA25 is much longer than LEA11 and COR15A, but shares a conserved central sequence domain with the other two proteins. We have created two truncated versions of LEA25 (2H and 4H) to elucidate the structural and functional significance of this domain. Light scattering and CD spectroscopy showed that all five proteins were largely unstructured and monomeric in dilute solution. They folded in the presence of increasing concentrations of trifluoroethanol and glycerol. Additional folding was observed in the presence of glycerol and membranes. Fourier transform infra red spectroscopy revealed an interaction of the LEA proteins with membranes in the dry state leading to a depression in the gel to liquid-crystalline phase transition temperature. Liposome stability assays revealed a cryoprotective function of the proteins. The C- and N-terminal extensions of LEA25 were important in cryoprotection, as the central domain itself (2H, 4H) only provided a low level of protection.

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“…Throughout the land plant lineage, water stress responses are coordinated by the plant growth regulator abscisic acid (ABA). ABA is synthesized in response to water deficit, and it elicits a range of responses to water stress, from the execution of stomatal closure to the induction of gene expression leading to the accumulation of protective compounds in desiccated tissues (typically disaccharide sugars and Late Embryogenesis Abundant [LEA] proteins required for the stabilization of cellular macromolecules and membranes in the dry state) (Koster and Leopold, 1988;Tunnacliffe and Wise, 2007;Buitink and Leprince, 2008;Shimizu et al, 2010;Clarke et al, 2015;Popova et al, 2015). The response to ABA is achieved through a highly conserved core signaling pathway, in which a family of receptor proteins (the PYRABACTIN RESISTANCE1 [PYR1]/PYR1-LIKE [PYL]/REGULATORY COMPONENTS OF ABA RECEPTORS [RCAR] proteins) bind ABA, which potentiates a molecular interaction with protein phosphatase 2C (PP2C) enzymes (e.g., those encoded by the ABSCISIC ACID INSENSITIVE1 [ABI1] and ABI2 genes).…”

Section: Introductionmentioning

confidence: 99%

Genetic analysis of Physcomitrella patens identifies ABSCISIC ACID NON-RESPONSIVE (ANR), a regulator of ABA responses unique to basal land plants and required for desiccation tolerance

et al. 2016

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The anatomically simple plants that first colonized land must have acquired molecular and biochemical adaptations to drought stress. Abscisic acid (ABA) coordinates responses leading to desiccation tolerance in all land plants. We identified ABA nonresponsive mutants in the model bryophyte Physcomitrella patens and genotyped a segregating population to map and identify the ABA NON-RESPONSIVE (ANR) gene encoding a modular protein kinase comprising an N-terminal PAS domain, a central EDR domain, and a C-terminal MAPKKK-like domain. anr mutants fail to accumulate dehydration tolerance-associated gene products in response to drought, ABA, or osmotic stress and do not acquire ABA-dependent desiccation tolerance. The crystal structure of the PAS domain, determined to 1.7-Å resolution, shows a conserved PAS-fold that dimerizes through a weak dimerization interface. Targeted mutagenesis of a conserved tryptophan residue within the PAS domain generates plants with ABA nonresponsive growth and strongly attenuated ABA-responsive gene expression, whereas deleting this domain retains a fully ABA-responsive phenotype. ANR orthologs are found in early-diverging land plant lineages and aquatic algae but are absent from more recently diverged vascular plants. We propose that ANR genes represent an ancestral adaptation that enabled drought stress survival of the first terrestrial colonizers but were lost during land plant evolution.

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Structural and Functional Insights into the Cryoprotection of Membranes by the Intrinsically Disordered Dehydrins

Cited by 47 publications

References 69 publications

Membrane-induced folding of the plant-stress protein Lti30.

Membrane-induced folding of the plant-stress protein Lti30.

Folding of intrinsically disordered plant LEA proteins is driven by glycerol‐induced crowding and the presence of membranes

Genetic analysis of Physcomitrella patens identifies ABSCISIC ACID NON-RESPONSIVE (ANR), a regulator of ABA responses unique to basal land plants and required for desiccation tolerance

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