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Antagonistic peptide technology for functional dissection of CLE peptides revisited.

Czyzewicz N, Wildhagen M, Cattaneo P, Stahl Y, Pinto KG, Aalen RB, Butenko MA, Simon R, Hardtke CS, De Smet I - J. Exp. Bot. (2015)

Bottom Line: Based on the analyses, it was concluded that the antagonistic peptide approach is not the ultimate means to overcome redundancy or lack of loss-of-function lines.However, information collected using antagonistic peptide approaches (in the broad sense) can be very useful, but these approaches do not work in all cases and require a deep insight on the interaction between the ligand and its receptor to be successful.This, as well as peptide ligand structure considerations, should be taken into account before ordering a wide range of synthetic peptide variants and/or generating transgenic plants.

View Article: PubMed Central - PubMed

Affiliation: Division of Plant and Crop Sciences, School of Biosciences, University of Nottingham, Loughborough LE12 5RD, UK.

No MeSH data available.


Alignment of CLE peptides used in Song et al. (2013). Conserved glycine (G) at position six is indicated with a blue arrowhead. (This figure is available in colour at JXB online.)
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Figure 1: Alignment of CLE peptides used in Song et al. (2013). Conserved glycine (G) at position six is indicated with a blue arrowhead. (This figure is available in colour at JXB online.)

Mentions: To interfere with and unravel endogenous peptide function, antagonistic peptides—such as mutant peptide variants, chemically modified peptides or peptide-like molecules that can affect peptide ligand–receptor (kinase) pathways are an important tool. In this context, structure-function/activity analyses can provide useful information on peptide residues critical for function. With respect to CLE peptides, such analyses were used to test, for example, suppression of nodulation capability in soybean (Glycine max) roots of the nodulation-controlling RHIZOBIA-INDUCED CLE1 (GmRIC1) (Reid et al., 2013) or regulation of primary and lateral root growth of various CLE peptides (Czyzewicz et al., 2015; Kondo et al., 2008). Recently, this approach was used to develop a promising new tool, referred to as antagonistic peptide technology, for functional dissection of CLE peptides (Song et al., 2013). Based on transgenic plants carrying CLV3 variants where each of the 12 residues in the core CLE motif were one by one replaced by alanine (Ala), it was shown that the glycine (Gly) to Ala substitution at position six gave a weak clv3 phenotype. Subsequently, replacing this highly conserved Gly residue with other amino acids revealed that a Gly to threonine (Thr) produced a phenotype most similar to clv3 mutants. This was further tested using synthetic CLV3 peptide with the Gly to Thr substitution (CLV3p6Thr), which was also able to produce—although less effective—the clv3 mutant phenotype, and which could compete with wild-type synthetic CLV3 peptide (CLV3p). These exciting observations suggested that the CLV3p6Thr variant could act as an antagonistic peptide. Specifically, a loss-of-function phenotype is suggested to be obtained through competitive inhibition, namely the peptide is proposed to be able to bind to the native receptor, but unable to activate it, since a functionally critical amino acid is mutated. Probably the CLV3p6Thr variant has compromised peptide flexibility leading to stronger interaction with corresponding receptors and to disrupted downstream signal transduction. Taken together, such antagonistic peptides would provide a powerful tool for the functional dissection of CLEs in plants, and might also have the potential to be used for other plant peptides. Based on this assumption and the conserved nature of the Gly at position six (Fig. 1), this technology was applied to CLE8 (giving rise to embryo-lethal phenotype) and CLE22 (giving rise to short root phenotype) (Song et al., 2013).


Antagonistic peptide technology for functional dissection of CLE peptides revisited.

Czyzewicz N, Wildhagen M, Cattaneo P, Stahl Y, Pinto KG, Aalen RB, Butenko MA, Simon R, Hardtke CS, De Smet I - J. Exp. Bot. (2015)

Alignment of CLE peptides used in Song et al. (2013). Conserved glycine (G) at position six is indicated with a blue arrowhead. (This figure is available in colour at JXB online.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4526918&req=5

Figure 1: Alignment of CLE peptides used in Song et al. (2013). Conserved glycine (G) at position six is indicated with a blue arrowhead. (This figure is available in colour at JXB online.)
Mentions: To interfere with and unravel endogenous peptide function, antagonistic peptides—such as mutant peptide variants, chemically modified peptides or peptide-like molecules that can affect peptide ligand–receptor (kinase) pathways are an important tool. In this context, structure-function/activity analyses can provide useful information on peptide residues critical for function. With respect to CLE peptides, such analyses were used to test, for example, suppression of nodulation capability in soybean (Glycine max) roots of the nodulation-controlling RHIZOBIA-INDUCED CLE1 (GmRIC1) (Reid et al., 2013) or regulation of primary and lateral root growth of various CLE peptides (Czyzewicz et al., 2015; Kondo et al., 2008). Recently, this approach was used to develop a promising new tool, referred to as antagonistic peptide technology, for functional dissection of CLE peptides (Song et al., 2013). Based on transgenic plants carrying CLV3 variants where each of the 12 residues in the core CLE motif were one by one replaced by alanine (Ala), it was shown that the glycine (Gly) to Ala substitution at position six gave a weak clv3 phenotype. Subsequently, replacing this highly conserved Gly residue with other amino acids revealed that a Gly to threonine (Thr) produced a phenotype most similar to clv3 mutants. This was further tested using synthetic CLV3 peptide with the Gly to Thr substitution (CLV3p6Thr), which was also able to produce—although less effective—the clv3 mutant phenotype, and which could compete with wild-type synthetic CLV3 peptide (CLV3p). These exciting observations suggested that the CLV3p6Thr variant could act as an antagonistic peptide. Specifically, a loss-of-function phenotype is suggested to be obtained through competitive inhibition, namely the peptide is proposed to be able to bind to the native receptor, but unable to activate it, since a functionally critical amino acid is mutated. Probably the CLV3p6Thr variant has compromised peptide flexibility leading to stronger interaction with corresponding receptors and to disrupted downstream signal transduction. Taken together, such antagonistic peptides would provide a powerful tool for the functional dissection of CLEs in plants, and might also have the potential to be used for other plant peptides. Based on this assumption and the conserved nature of the Gly at position six (Fig. 1), this technology was applied to CLE8 (giving rise to embryo-lethal phenotype) and CLE22 (giving rise to short root phenotype) (Song et al., 2013).

Bottom Line: Based on the analyses, it was concluded that the antagonistic peptide approach is not the ultimate means to overcome redundancy or lack of loss-of-function lines.However, information collected using antagonistic peptide approaches (in the broad sense) can be very useful, but these approaches do not work in all cases and require a deep insight on the interaction between the ligand and its receptor to be successful.This, as well as peptide ligand structure considerations, should be taken into account before ordering a wide range of synthetic peptide variants and/or generating transgenic plants.

View Article: PubMed Central - PubMed

Affiliation: Division of Plant and Crop Sciences, School of Biosciences, University of Nottingham, Loughborough LE12 5RD, UK.

No MeSH data available.