PLANT PHYSIOLOGY , Vol 106, Issue 2 567-574, Copyright © 1994 by American Society of Plant Biologists

MOLECULAR BIOLOGY AND GENE REGULATION

Inhibitors of Protein Phosphatases 1 and 2A Block the Sugar-Inducible Gene Expression in Plants

S. Takeda, S. Mano, Ma. Ohto and K. Nakamura
Laboratory of Biochemistry, School of Agriculture, Nagoya University, Chikusa, Nagoya 464-01, Japan

Genes coding for two major proteins of the tuberous root of sweet potato (Ipomoea batatas), namely, sporamin and [beta]-amylase, are inducible in leaves and petioles when they are supplied with high concentrations of sucrose or other metabolizable sugars, such as glucose and fructose, and the accumulation of a large amount of starch accompanies this induction. Three inhibitors of protein phosphatases 1 (PP1) and 2A (PP2A), namely, okadaic acid, microcystin-LR, and calyculin A, strongly inhibited the sucrose-inducible accumulation of mRNAs for sporamin, [beta]-amylase, and the small subunit of ADP-glucose pyrophosphorylase in petioles. However, these inhibitors did not have any major effect on the steady-state levels of mRNAs for catalase and glyceraldehyde-3-phosphate dehydrogenase, and the sucrose-inducible increase in the level of sucrose synthase mRNA was enhanced by okadaic acid. Inhibitors of PP1 and PP2A also inhibited sucrose-inducible expression of a fusion gene, consisting of the promoter of the sweet potato gene for [beta]-amylase and the coding sequence for [beta]-glucuronidase (GUS), in leaves of transgenic tobacco (Nicotiana tabacum). The inhibition was not due to inhibition of uptake and cleavage of sucrose, since okadaic acid also inhibited induction of the fusion gene by glucose or fructose. Addition of okadaic acid to leaves that had been treated with sucrose for 6 h inhibited further increases in GUS activity. These results suggest that the continuous dephosphorylation of proteins is required in the transduction of carbohydrate metabolic signals to the transcriptional activation of at least some sugar-inducible genes in plant.

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