Abstract

Tap46, a regulatory subunit of protein phosphatase 2A (PP2A), plays an essential role in plant growth and development through a functional link with the Target of Rapamycin (TOR) signalling pathway. Here, we have characterized the molecular mechanisms behind a gain-of-function phenotype of Tap46 and its relationship with TOR to gain further insights into Tap46 function in plants. Constitutive overexpression of Tap46 in Arabidopsis resulted in overall growth stimulation with enlarged organs, such as leaves and siliques. Kinematic analysis of leaf growth revealed that increased cell size was mainly responsible for the leaf enlargement. Tap46 overexpression also enhanced seed size and viability under accelerated ageing conditions. Enhanced plant growth was also observed in dexamethasone (DEX)-inducible Tap46 overexpression Arabidopsis lines, accompanied by increased cellular activities of nitrate-assimilating enzymes. DEX-induced Tap46 overexpression and Tap46 RNAi resulted in increased and decreased phosphorylation of S6 kinase (S6K), respectively, which is a sensitive indicator of endogenous TOR activity, and Tap46 interacted with S6K in planta based on bimolecular fluorescence complementation and co-immunoprecipitation. Furthermore, inactivation of TOR by estradiol-inducible RNAi or rapamycin treatment decreased Tap46 protein levels, but increased PP2A catalytic subunit levels. Real-time quantitative PCR analysis revealed that Tap46 overexpression induced transcriptional modulation of genes involved in nitrogen metabolism, ribosome biogenesis, and lignin biosynthesis. These findings suggest that Tap46 modulates plant growth as a positive effector of the TOR signalling pathway and Tap46/PP2Ac protein abundance is regulated by TOR activity.

Highlights

  • Target of Rapamycin (TOR) is a Ser/Thr protein kinase that is evolutionarily conserved among all eukaryotes

  • DEX-induced Tap46 overexpression and Tap46 RNAi resulted in increased and decreased phosphorylation of S6 kinase (S6K), respectively, which is a sensitive indicator of endogenous TOR activity, and Tap46 interacted with S6K in planta based on bimolecular fluorescence complementation and co-immunoprecipitation

  • These findings suggest that Tap46 modulates plant growth as a positive effector of the TOR signalling pathway and Tap46/ PP2A catalytic subunit (PP2Ac) protein abundance is regulated by TOR activity

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Summary

Introduction

Target of Rapamycin (TOR) is a Ser/Thr protein kinase that is evolutionarily conserved among all eukaryotes. The TOR signalling pathway integrates diverse signals, such as nutrients, energy, growth factors, and environmental conditions, to regulate cell growth and metabolism in yeast, plants, and mammals (Hay and Sonenberg, 2004; Wullschleger et al, 2006; Bögre et al, 2013; Xiong and Sheen, 2013). TOR kinase forms two distinct multiprotein complexes in yeast and mammals, named TOR complex 1 (TORC1) and TOR complex 2 (TORC2), which contain both conserved and distinct protein components.

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