Wheat cysteine proteases triticain α, β and γ exhibit mutually distinct responses to gibberellin in germinating seeds

Kiyosaki, Toshihiro; Asakura, Tomiko; Matsumoto, Ichiro; Tamura, Tomoko; Terauchi, Kaede; Funaki, Junko; Kuroda, Masaharu; Misaka, Takumi; Abe, Keiko

doi:10.1016/j.jplph.2008.02.006

Cited by 31 publications

(23 citation statements)

References 22 publications

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“…The expression of other known cereal GA response genes, including GAMYB, a transcription regulator of GA signaling in aleurone (Gubler et al, 1995(Gubler et al, , 1999Murray et al, 2006), a cell wall-degrading enzyme, (1,3;1,4)-b-glucanase (Mundy and Fincher, 1986), and two proteases, triticain-a and triticain-g (Kiyosaki et al, 2009), was also determined in mature aleurone. In comparison with the expression on control medium, the expression of (1,3;1,4)-b-glucanase was induced 5-fold by GA, triticain-a was induced 14-fold, triticain-g was induced 32-fold, and GAMYB was induced 3-fold (Fig.…”

Section: Expression Of A-amylase Genes and Ga Response Genes In Maturmentioning

confidence: 99%

Genetic, Hormonal, and Physiological Analysis of Late Maturity α-Amylase in Wheat

et al. 2013

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Late maturity a-amylase (LMA) is a genetic defect that is commonly found in bread wheat (Triticum aestivum) cultivars and can result in commercially unacceptably high levels of a-amylase in harvest-ripe grain in the absence of rain or preharvest sprouting. This defect represents a serious problem for wheat farmers, and apart from the circumstantial evidence that gibberellins are somehow involved in the expression of LMA, the mechanisms or genes underlying LMA are unknown. In this work, we use a doubled haploid population segregating for constitutive LMA to physiologically analyze the appearance of LMA during grain development and to profile the transcriptomic and hormonal changes associated with this phenomenon. Our results show that LMA is a consequence of a very narrow and transitory peak of expression of genes encoding high-isoelectric point a-amylase during grain development and that the LMA phenotype seems to be a partial or incomplete gibberellin response emerging from a strongly altered hormonal environment.

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Section: Expression Of A-amylase Genes and Ga Response Genes In Maturmentioning

confidence: 99%

Genetic, Hormonal, and Physiological Analysis of Late Maturity α-Amylase in Wheat

et al. 2013

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“…These endopeptidases are synthesised de novo in the scutellum and aleurone layer in response to gibberellin signalling (Fincher 1989;Holwerda et al 1990;Mikkonen et al 1996;Zhang and Jones 1996). It was confirmed by evaluating their transcript and activity level in rice (Watanabe et al 1991), wheat (Kiyosaki et al 2007(Kiyosaki et al , 2009Shi and Xu 2009) and barley (Mikkonen et al 1996;Martinez et al 2009). In our previous study the activity of seven CEPs, EP2-EP8, and one aspartyl proteinase, EP1, hydrolysing wheat gliadin, was observed in the endosperm of triticale grains (Prabucka and Bielawski 2004).…”

Section: Discussionmentioning

confidence: 87%

The participation of phytocystatin TrcC-4 in the activity regulation of EP8, the main prolamin degrading cysteine endopeptidase in triticale seeds

et al. 2012

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Phytocystatins (PCs) are protein inhibitors of endogenous plant endopeptidases and exogenous pathogen proteinases. We have previously described the protein inhibitor TrcC-4, which is probably involved in the control of protein degradation during triticale seeds germination. The occurrence of the LARFAVXEHN motif supports the TrcC-4 designation as a PC. In this paper TrcC-4 was expressed in Escherichia coli using the pET28 expression vector. TrcC-4(69His) was purified by affinity chromatography with a single step of purification. Western blot analysis showed the presence of TrcC-4 in both developing and germinating triticale seeds. TrcC-4 protein level was higher both in scutellum of germinating seeds and in developing grains of the triticale cultivar more resistant to pre-harvest sprouting (Zorro) than in a less resistant one (Disco). Furthermore it was demonstrated that the activity of EP8, cysteine endopeptidase responsible for the mass hydrolysis of prolamin during germination, is inhibited by TrcC-4(69His), as confirmed by native PAGE with gliadin as a substrate. These results suggest that phytocystatin TrcC-4 controls the activity of cysteine endopeptidases involved in germination and, thus, is potentially involved in pre-harvest sprouting.

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“…Expression analysis of Z-Phe-Arg-MCA cleaving activity following germination Cotyledons (20 g) from germinating embryo (8,24,36,48,72, 144 h after imbibition) were homogenized with 100 ml of 20 mM acetate buffer (pH 6.5) containing 1 mM b-mercaptoethanol and 0.5 mM EDTA (buffer A). Following centrifugation at 20,000 Â g for 10 min, the supernatant was fractionated by ammonium sulfate precipitation (60% saturation) and then dialysed against buffer A.…”

Section: Enzyme Assaymentioning

confidence: 99%

“…Glucagon, ACTH (1 24) and somatostatin-28 were digested, and the cleavage sites were identified. Five glucagon-derived 24 . These results indicated that the cleavage specificities of DRCP26, CBCP and RD21 differed.…”

Section: Purification and Characterization Of Drcp26mentioning

confidence: 99%

Enzymatic characterization of germination-specific cysteine protease-1 expressed transiently in cotyledons during the early phase of germination

Tsuji¹,

Tsukamoto²,

Iwamoto³

et al. 2012

Journal of Biochemistry

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Papain-like cysteine protease activity that shows a unique transient expression profile in cotyledons of daikon radish during germination was detected. The enzyme showed a distinct elution pattern on DEAE-cellulose compared with cathepsin B-like and Responsive to dessication-21 cysteine protease. Although this activity was not detected in seed prior to imbibition, the activity increased markedly and reached a maximum at 2 days after imbibition and then decreased rapidly and completely disappeared after 5 days. Using cystatin-Sepharose, the 26 kDa cysteine protease (DRCP26) was isolated from cotyledons at 2 days after imbibition. The deduced amino acid sequence from the cDNA nucleotide sequence indicated that DRCP26 is an orthologue of Arabidopsis unidentified protein, germination-specific cysteine protease-1, belonging to the C1 family of cysteine protease predicted from genetic information. In an effort to characterize the enzymatic properties of DRCP26, the enzyme was purified to homogeneity from cotyledons at 48 h after imbibition. The best synthetic substrate for the enzyme was carbobenzoxy-Phe-Arg-4-methylcoumaryl-7-amide. All model peptides were digested to small peptides by the enzyme, suggesting that DRCP26 possesses broad cleavage specificity. These results indicated that DRCP26 plays a role in the mobilization of storage proteins in the early phase of seed germination.

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Wheat cysteine proteases triticain α, β and γ exhibit mutually distinct responses to gibberellin in germinating seeds

Cited by 31 publications

References 22 publications

Genetic, Hormonal, and Physiological Analysis of Late Maturity α-Amylase in Wheat

Genetic, Hormonal, and Physiological Analysis of Late Maturity α-Amylase in Wheat

The participation of phytocystatin TrcC-4 in the activity regulation of EP8, the main prolamin degrading cysteine endopeptidase in triticale seeds

Enzymatic characterization of germination-specific cysteine protease-1 expressed transiently in cotyledons during the early phase of germination

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