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GrainGenes Reference Report: PMB-46-361

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Reference	PMB-46-361
Title	Characterization of two thioredoxins h with predominant localization in the nucleus of aleurone and scutellum cells of germinating wheat seeds
Journal	Plant Molecular Biology
Year	2001
Volume	46
Pages	361-371
Author	Serrato AJ Crespo JL Florencio FJ Cejudo FJ
Abstract	Summary: Two full-length cDNA clones, designated TrxhA and TrxhB, encoding different but very similar thioredoxin h polypeptides were isolated from wheat (Triticum aestivum cv. Chinese Spring) aleurone cells. The deduced proteins show a high similarity to each other and to thioredoxin h from other sources, in particular from T. aestivum and T. durum. One of them, TRXhA, was expressed in E. coli as a His-tagged polypeptide and used to raise polyclonal antibodies by immunization of rabbits. These antibodies identified a single band (ca. 13.5 kDa) in western blot analysis of protein extracts from all wheat organs analyzed. TRXhA and TRXhB when expressed in E. coli as intact polypeptides showed indistinguishable electrophoretic mobility, which corresponded to the 13.5 kDa polypeptide detected in wheat protein extracts. The amount of thioredoxin h transcripts increased in scutellum and aleurone cells during germination but GA3 did not exert any stimulatory effect on thioredoxin h expression. Although northern blot analysis detected a single band, competitive RT-PCR showed that this band is due to the accumulation of both TrxhA and TrxhB mRNAs. These results suggest that the single band detected in western blots is due to the presence of at least two thioredoxin h polypeptides. Immunolocalization experiments confirmed the high content of thioredoxins h in scutellum and aleurone cells, and showed a low content in the starchy endosperm of germinating grains. Interestingly, though these proteins are evenly distributed in the cytosol, the highest levels of thioredoxins h were detected in the nucleus, both in aleurone and scutellum cells
Keyword	[ Hide all but 1 of 46 ] acid aleurone aleurone cells antibody barley aleurone cdna cdna clone cell characterization chinese chinese Spring clone cytosol death durum endosperm full-length cdna germinating wheat germination gibberellic acid grains localization mobility mrna northern nuclear localization nucleus plant science polypeptide protein reduction saccharomyces cerevisiae scutellum seed single spring starchy endosperm system thioredoxin thioredoxin h transcript triticum triticum aestivum western blot analysis wheat protein wheat seed

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