《《+PlantCell-SIZ1-Sumoylation-ICE1-CBF3-2016_81302120》.pdf

The Plant Cell, Vol. 19: 1403–1414, April 2007, ª 2007 American Society of Plant Biologists SIZ1-Mediated Sumoylation of ICE1 Controls CBF3/DREB1A Expression and Freezing Tolerance in Arabidopsis W OA Kenji Miura,a Jing Bo Jin,a Jiyoung Lee,a Chan Yul Yoo,a Vicki Stirm,a Tomoko Miura,a Edward N. Ashworth,b Ray A. Bressan,a Dae-Jin Yun,c,1 and Paul M. Hasegawaa,1 a Center for Plant Environmental Stress Physiology, Purdue University, West Lafayette, Indiana 47907-2010 b College of Natural Sciences, Forestry and Agriculture, University of Maine, Orono, Maine 04469-5782 c Division of Applied Life Science (BK21 Program), Plant Molecular Biology and Biotechnology Research Center, Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea SIZ1 is a SUMO E3 ligase that facilitates conjugation of SUMO to protein substrates. siz1-2 and siz1-3 T-DNA insertion alleles that caused freezing and chilling sensitivities were complemented genetically by expressing SIZ1, indicating that the SIZ1 is a controller of low temperature adaptation in plants. Cold-induced expression of CBF/DREB1, particularly of CBF3/ DREB1A, and of the regulon genes was repressed by siz1. siz1 did not affect expression of ICE1, which encodes a MYC transcription factor that is a controller of CBF3/DREB1A. A K393R substitution in ICE1 [ICE1(K393R)] blocked SIZ1-mediated sumoylation in vitro and in protoplasts identifying the K393 residue as the principal site of SUMO conjugation. SIZ1- dependent sumoylation of ICE1 in protoplasts was moderately induced by cold. Sumoylation of recombinant ICE1 reduced polyubiquitination of the protein in vitro. ICE1(K393R) expression in wild-type plants repressed cold-induced CBF3/DREB1A expression and increased freezing sensitivity. Furthermore, expression of ICE1(K393R) induced transcript accumulati