BLISTER-regulated vegetative growth is dependent on the protein kinase domain of ER stress modulator IRE1A in Arabidopsis thaliana.

Zheng-Hui Hong,Julia Anna Kleinmanns,Tao Qing,Jian-Xiang Liu,Daniel Schubert,Li-Jia Qu

doi:10.1371/journal.pgen.1008563

Abstract

The unfolded protein response (UPR) is required for protein homeostasis in the endoplasmic reticulum (ER) when plants are challenged by adverse environmental conditions. Inositol-requiring enzyme 1 (IRE1), the bifunctional protein kinase / ribonuclease, is an important UPR regulator in plants mediating cytoplasmic splicing of the mRNA encoding the transcription factor bZIP60. This activates the UPR signaling pathway and regulates canonical UPR genes. However, how the protein activity of IRE1 is controlled during plant growth and development is largely unknown. In the present study, we demonstrate that the nuclear and Golgi-localized protein BLISTER (BLI) negatively controls the activity of IRE1A/IRE1B under normal growth condition in Arabidopsis. Loss-of-function mutation of BLI results in chronic up-regulation of a set of both canonical UPR genes and non-canonical UPR downstream genes, leading to cell death and growth retardation. Genetic analysis indicates that BLI-regulated vegetative growth phenotype is dependent on IRE1A/IRE1B but not their canonical splicing target bZIP60. Genetic complementation with mutation analysis suggests that the D570/K572 residues in the ATP-binding pocket and N780 residue in the RNase domain of IRE1A are required for the activation of canonical UPR gene expression, in contrast, the D570/K572 residues and D590 residue in the protein kinase domain of IRE1A are important for the induction of non-canonical UPR downstream genes in the BLI mutant background, which correlates with the shoot growth phenotype. Hence, our results reveal the important role of IRE1A in plant growth and development, and BLI negatively controls IRE1A’s function under normal growth condition in plants.

Highlights

Protein folding in the endoplasmic reticulum (ER) is a fundamental process in eukaryotic cells
When unfolded or misfolded proteins are accumulated in the ER, a much conserved response, called the unfolded protein response (UPR), is elicited to lighten the load of unfolded proteins in the ER by bringing the protein-folding and degradation capacities into alignment with the protein folding demands
We report that BLISTER (BLI) negatively controls the Inositol-requiring enzyme 1 (IRE1)’s function under normal growth condition in Arabidopsis

Summary

Introduction

Protein folding in the ER is a fundamental process in eukaryotic cells. Protein folding demands on the secretory pathway escalate constantly during different developmental stages and environmental conditions while protein folding capacity is limited depending on ER chaperones, oxidoreductases, N-glycosylation etc. [1,2]. Like yeast and mammalian cells, plant cells are equipped with a set of membrane-associated transcription factors for sensing/transducing ER stress signals, the sequence similarities among these factors are modest [4,5,6,7,8,9,10,11,12,13,14]. Upon ER stress induced by chemicals or abiotic stresses such as heat stress, bZIP28 relocates from ER to Golgi, where it is subjected to proteolysis by Golgi-resident Site-2 Protease (S2P) This cleavage releases its cytoplasmic N-terminal region that contains a DNA-binding domain, a nuclear localization signal and a transcriptional activation domain for downstream stress responsive gene expression [4,15,16,17,18,19,20]. How IRE1 activity is controlled under normal growth conditions in plants is currently unknown

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