A conserved dibasic site is essential for correct processing of the peptide hormone AtRALF1 in <i>Arabidopsis thaliana</i>

Matos, Juliana L.; Fiori, Celso S; Silva-Filho, Márcio C.; Moura, Daniel S.

doi:10.1016/j.febslet.2008.08.025

Cited by 89 publications

(131 citation statements)

References 24 publications

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“…The peptide was cleaved at Met residues by cyanogen bromide, and the resulting mixture was analyzed by MS. Two fragments were identified with molecular masses of 1628 and 1831 Da. The latter corresponds to the middle part of the peptide (residues [9][10][11][12][13][14][15][16][17][18][19][20][21][22] containing the 'inner' Cys 9 -Cys 21 disulfide bond. The former corresponds to the two flanking parts (residues 1-8 and 23-28) held together by the 'outer' Cys 5 -Cys 25 disulfide bond.…”

Section: Resultsmentioning

confidence: 99%

Genes encoding 4‐Cys antimicrobial peptides in wheat Triticum kiharae Dorof. et Migush.: multimodular structural organization, instraspecific variability, distribution and role in defence

et al. 2013

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A novel family of antifungal peptides was discovered in the wheat Triticum kiharae Dorof. et Migusch. Two members of the family, designated Tk-AMP-X1 and Tk-AMP-X2, were completely sequenced and shown to belong to the a-hairpinin structural family of plant peptides with a characteristic C1XXXC2-X(n)-C3XXXC4 motif. The peptides inhibit the spore germination of several fungal pathogens in vitro. cDNA and gene cloning disclosed unique structure of genes encoding Tk-AMP-X peptides. They code for precursor proteins of unusual multimodular structure, consisting of a signal peptide, several a-hairpinin (4-Cys) peptide domains with a characteristic cysteine pattern separated by linkers and a C-terminal prodomain. Three types of precursor proteins, with five, six or seven 4-Cys peptide modules, were found in wheat. Among the predicted family members, several peptides previously isolated from T. kiharae seeds were identified. Genes encoding Tk-AMP-X precursors have no introns in the proteincoding regions and are upregulated by fungal pathogens and abiotic stress, providing conclusive evidence for their role in stress response. A combined PCR-based and bioinformatics approach was used to search for related genes in the plant kingdom. Homologous genes differing in the number of peptide modules were discovered in phylogenetically-related Triticum and Aegilops species, including polyploid wheat genome donors. Association of the Tk-AMP-X genes with A, B/G or D genomes of hexaploid wheat was demonstrated. Furthermore, Tk-AMP-X-related sequences were shown to be widespread in the Poaceae family among economically important crops, such as barley, rice and maize.

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Section: Resultsmentioning

confidence: 99%

Genes encoding 4‐Cys antimicrobial peptides in wheat Triticum kiharae Dorof. et Migush.: multimodular structural organization, instraspecific variability, distribution and role in defence

et al. 2013

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“…The loss-of-function gene mutations SDD1 (AtSBT1.2) and ALE1 (AtSBT2.4) lead to abnormal stomatal density and leaf shape, respectively. These results indicate that some subtilases may play a role in developmental processes through the generation of peptide signals (von Groll et al, 2002;Matos et al, 2008). Overexpression of another subtilase (AtSBT5.4) produces a clavata-like phenotype with fasciated inflorescence stems and compounded terminal buds.…”

Section: Introductionmentioning

confidence: 96%

Molecular cloning and characterization of a subtilisin-like protease from Arabidopsis thaliana

et al. 2015

Genet. Mol. Res.

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ABSTRACT. The Arabidopsis thaliana genome encodes 56 subtilisin-like serine proteases (subtilases). In order to evaluate the protease activity of a previously uncharacterized subtilase, designated as AtSBT1.9, we cloned its full-length cDNA from A. thaliana seedlings. An AtSBT1.9 mature peptide coding sequence was inserted into the bacterial expression vector, pMALc2x, and the recombinant vector was transformed into Escherichia coli BL21 (DE3). The recombinant AtSBT1.9 tagged by maltose binding protein (MBP) was induced as a 117.5-kDa protein in the soluble form in E. coli BL21 (DE3). MBP-AtSBT1.9 was expressed at a level of 11% (w/w) of the bacterial total protein. Protein purification using Amylose Resin revealed a recombinant AtSBT1.9 protease activity of 9.23 U/mg protein at pH 7 and 25°C. Maximal activity occurred over a broad pH (7-8) and temperature (25°-42°C) optimal range. Validation of AtSBT1.9 protease activity would help in characterizing its in vivo function in A. thaliana.

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“…[19][20][21] RALF precursors possess a conserved dibasic site upstream to the active peptide, and recent studies have shown that this site is required for pro-peptide processing and activity of both AtRALF1 and AtRALF23. 22,23 In the case of tomato pollen SlRALF, mature peptide was only detected in the medium in an in vitro pollen germination system. 18 These results are also consistent with localization of a Nicotiana benthamiana RALF fused to GFP, which localized first to the ER and later to the cell wall in N. benthamiana leaf cells.…”

Section: Ralf Processing and Localizationmentioning

confidence: 99%

“…Transgenic studies in Arabidopsis showed that overexpression of either AtRALF1 or AtRALF23 resulted in semi-dwarf plants. 22,23 In Medicago trunculata, a root-expressed RALF gene (MtRALFL1) was discovered as being upregulated by nodulation factors. 35 Overexpression of the MtRALFL1 gene in transgenic plants resulted in a reduction in nodules and an increase in aborted infection threads.…”

Section: Possible Mechanisms Of Ralf Actionmentioning

confidence: 99%

RALFs

Bedinger¹,

Pearce²,

Covey³

2010

Plant Signaling & Behavior

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The Discovery of RALFDuring a search for systemins in tobacco, an unrelated peptide factor was discovered that alkalinizes suspension culture medium even more rapidly than systemins. 13 This factor was named RALF, for Rapid ALkalinization Factor. RALF peptide was first purified from tobacco leaves using the suspension cell alkalinization assay, and an N-terminal peptide sequence was obtained. This sequence allowed the identification of a full length tobacco cDNA indicating that the RALF gene encodes a 115 aa secreted preproprotein that is processed to release a 49 aa active peptide from the C-terminus. Highly conserved active RALF peptides were also isolated from tomato and alfalfa cells, and a synthetic peptide based on the tomato sequence was produced. Both native and oxidized synthetic peptides were able to produce alkalinization responses at nanomolar concentrations, but alkylation of the reduced form of the peptide (which prevents formation of disulfide bridges) inactivated the synthetic peptide. A role for RALFs in defense was considered, since both systemins and RALFs induce alkalinization of medium and MAP kinase activation. However, application of RALF peptide to tobacco leaves did not induce a defense response as detectable by the induction of tobacco trypsin inhibitor. Studies using poplar-derived RALFs

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A conserved dibasic site is essential for correct processing of the peptide hormone AtRALF1 in Arabidopsis thaliana

Cited by 89 publications

References 24 publications

Genes encoding 4‐Cys antimicrobial peptides in wheat Triticum kiharae Dorof. et Migush.: multimodular structural organization, instraspecific variability, distribution and role in defence

Genes encoding 4‐Cys antimicrobial peptides in wheat Triticum kiharae Dorof. et Migush.: multimodular structural organization, instraspecific variability, distribution and role in defence

Molecular cloning and characterization of a subtilisin-like protease from Arabidopsis thaliana

RALFs

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