Elicitin–membrane interaction is driven by a positive charge on the protein surface: Role of Lys13 residue in lipids loading and resistance induction

Plešková, Veronika; Kašparovský, Tomáš; Obořil, Michal; Ptáčková, Nikola; Chaloupková, Radka; Dokládal, Ladislav; Damborský, Jiřı́; Lochman, Jan

doi:10.1016/j.plaphy.2011.01.008

Cited by 15 publications

(15 citation statements)

References 45 publications

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“…So, it is obvious that the activity of elicitins is more dependent on the presence of specific residues than on the ω-loop structure; the most probable candidates being lysine residues in helices A and D of basic elicitins (Boissy et al , 1996). This assumption is further supported by the observed correlation between necrotic index and p I (Pernollet et al , 1993) and by a clear impact of the Lys13Val mutation in helix A on induction of a defence response in tobacco plants (Odonohue et al , 1995; Pleskova et al , 2011). …”

Section: Discussionmentioning

confidence: 72%

“…Previously, it has been shown that elicitins are able to bind and transfer sterols and phospholipids (through their fatty acid chains), for example, the SCP-2 protein from rat liver has similar binding and transfer properties for sterols and phospholipids (Mikes et al , 1998; Pleskova et al , 2011). Data from sterol (specifically, DHE) binding assays were in agreement with expectations based on the structural data of the DHE-cryptogein and fatty acid–cryptogein complexes reported previously (Dobeš et al , 2004).…”

Section: Discussionmentioning

confidence: 99%

“…The primers used for mutagenesis are given in Supplementary Table S2 at JXB online. The product was confirmed to have the expected DNA sequence, then the vector was used to transform Pichia pastoris and expression of the protein was induced in a Biostat B-DCU (Sartorius, Germany) fermentor using previously described protocol (Wood and Komives, 1999), with a methanol induction phase of 48 h. The expressed protein was purified by ultrafiltration and Fast Protein Liquid Chromatography using a Source S15 column as described before (Pleskova et al , 2011). …”

Section: Methodsmentioning

confidence: 99%

“…Unilamellar donor vesicles containing NBD-PC, phosphatidylserine (PS) and cholesterol and acceptor vesicles (containing PC, PS, and cholesterol) were prepared as described previously (Pleskova et al , 2011). Elicitin-induced phosphatidylcholine exchanges were measured by adding NBD-PC/PS (with 1.5 μM NBD-PC) donor vesicles to 2 ml measuring buffer (10 mM MES pH 7.0), containing PC/PS (with 3 μM PC) unilamellar acceptor vesicles, then recording the fluorescence of NBD-PC in both the absence (to determine spontaneous transfer rates) and presence of the elicitins.…”

Section: Methodsmentioning

confidence: 99%

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Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance

Dokládal

Obořil

Stejskal

et al. 2012

Journal of Experimental Botany

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Cryptogein is a proteinaceous elicitor secreted by Phytophthora cryptogea that can induce resistance to P. parasitica in tobacco plants. On the basis of previous computer modelling experiments, by site-directed mutagenesis a series of cryptogein variants was prepared with altered abilities to bind sterols, phospholipids or both. The sterol binding and phospholipid transfer activities corresponded well with the previously reported structural data. Induction of the synthesis of reactive oxygen species (ROS) in tobacco cells in suspension and proteomic analysis of intercellular fluid changes in tobacco leaves triggered by these mutant proteins were not proportional to their ability to bind or transfer sterols and phospholipids. However, changes in the intercellular proteome corresponded to transcription levels of defence genes and resistance to P. parasitica and structure-prediction of mutants did not reveal any significant changes in protein structure. These results suggest, contrary to previous proposals, that the sterol-binding ability of cryptogein and its mutants, and the associated conformational change in the ω-loop, might not be principal factors in either ROS production or resistance induction. Nevertheless, the results support the importance of the ω-loop for the interaction of the protein with the high affinity binding site on the plasma membrane.

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Section: Discussionmentioning

confidence: 72%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance

Dokládal

Obořil

Stejskal

et al. 2012

Journal of Experimental Botany

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“…Of the 12 B. lactucae protein models with elicitin domain matches, nine contained at least six cysteines in the predicted mature peptide. Phytophthora elicitins trigger a hypersensitive response in host plants, an effect that is linked to sterol binding and transporting activity for which a surface‐exposed polar residue is important (Hirasawa et al ., 2004; Mikes et al ., 1997, 1998; Plešková et al ., 2011; Ricci et al ., 1989; Vauthrin et al ., 1999). Elicitins may play a role in the uptake of sterols by oomycetes that do not synthesize their own, as the presence of elicitins appears to be correlated with the loss of the sterol biosynthetic pathway (Gaulin et al ., 2010; Jiang et al ., 2006; Vauthrin et al ., 1999).…”

Section: Resultsmentioning

confidence: 99%

Effector identification in the lettuce downy mildew Bremia lactucae by massively parallel transcriptome sequencing

Stassen

Seidl

Vergeer

et al. 2012

Molecular Plant Pathology

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Lettuce downy mildew (Bremia lactucae) is a rapidly adapting oomycete pathogen affecting commercial lettuce cultivation. Oomycetes are known to use a diverse arsenal of secreted proteins (effectors) to manipulate their hosts. Two classes of effector are known to be translocated by the host: the RXLRs and Crinklers. To gain insight into the repertoire of effectors used by B. lactucae to manipulate its host, we performed massively parallel sequencing of cDNA derived from B. lactucae spores and infected lettuce (Lactuca sativa) seedlings. From over 2.3 million 454 GS FLX reads, 59 618 contigs were assembled representing both plant and pathogen transcripts. Of these, 19 663 contigs were determined to be of B. lactucae origin as they matched pathogen genome sequences (SOLiD) that were obtained from >270 million reads of spore-derived genomic DNA. After correction of cDNA sequencing errors with SOLiD data, translation into protein models and filtering, 16 372 protein models remained, 1023 of which were predicted to be secreted. This secretome included elicitins, necrosis and ethylene-inducing peptide 1-like proteins, glucanase inhibitors and lectins, and was enriched in cysteine-rich proteins. Candidate host-translocated effectors included 78 protein models with RXLR effector features. In addition, we found indications for an unknown number of Crinkler-like sequences. Similarity clustering of secreted proteins revealed additional effector candidates. We provide a first look at the transcriptome of B. lactucae and its encoded effector arsenal.

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Differential induction of oxylipin pathway in potato and tobacco cells by bacterial and oomycete elicitors

Saubeau¹,

Goulitquer

Barloy

et al. 2013

Plant Cell Rep

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Potato and tobacco cells are differentially suited to study oxylipin pathway and elicitor-induced responses. Synthesis of oxylipins via the lipoxygenase (LOX) pathway provides plant cells with an important class of signaling molecules, related to plant stress responses and innate immunity. The aim of this study was to evaluate the induction of LOX pathway in tobacco and potato cells induced by a concentrated culture filtrate (CCF) from Phytophthora infestans and lipopolysaccharide (LPS) from Pectobacterium atrosepticum. Oxylipin activation was evaluated by the measurement of LOX activity and metabolite quantification. The basal levels of oxylipins and fatty acids showed that potato cells contained higher amounts of linoleic (LA), linolenic (LnA) and stearic acids than tobacco cells. The major oxylipin in potato cells, 9(S),10(S),11(R)-trihydroxy-12(Z),15(Z)-octadecadienoic acid (9,10,11-THOD), was not detected in tobacco cells. CCF induced a sharp increase of LA and LnA at 8 h in tobacco cells. In contrast they decreased in potato cells. In CCF-treated tobacco cells, colneleic acid increased up to 24 h, colnelenic acid and 9(S)-hydroxyoctadecatrienoic acid (9(S)-HOT) increased up to 16 h. In potato cells, only colneleic acid increased slightly until 16 h. A differential induction of LOX activity was measured in both cells treated by CCF. With LPS treatment, only 9,10,11-THOD accumulation was significantly induced at 16 h in potato cells. Fatty acids were constant in tobacco but decreased in potato cells over the studied time period. These results showed that the two elicitors were differently perceived by the two Solanaceae and that oxylipin pathway is strongly induced in tobacco with the CCF. They also revealed that elicitor-induced responses depended on both cell culture and elicitor.

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Elicitin–membrane interaction is driven by a positive charge on the protein surface: Role of Lys13 residue in lipids loading and resistance induction

Cited by 15 publications

References 45 publications

Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance

Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance

Effector identification in the lettuce downy mildew Bremia lactucae by massively parallel transcriptome sequencing

Differential induction of oxylipin pathway in potato and tobacco cells by bacterial and oomycete elicitors

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