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Published on February 11, 2009; 10.1104/pp.108.133744

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Received December 7, 2008
Accepted February 10, 2009

Molecular and genetic evidence for the key role of AtCaM3 in heat-shock signal transduction in Arabidopsis thaliana

Wei Zhang , Ren-Gang Zhou , Ying-Jie Gao , Shu-Zhi Zheng , Peng Xu , Su-Qiao Zhang , and Da-Ye Sun *

Institute of Molecular Cell Biology, Hebei Normal University, Shijiazhuang 050016, China; Institute of Genetics and Physiology, Hebei Academy of Agricultural Sciences, Shijiazhuang 050051, China; Center for Agricultural Resources Research Institute of Genetics and Developmental, Shijiazhuang 050021, China

* Corresponding author; email: dayesun{at}gmail.com.

Heat shock (HS) is a common form of stress suffered by plants. It has been proposed that calmodulin (CaM) is involved in HS signal transduction, but direct evidence has been lacking. To investigate the potential regulatory function of CaM in the HS signal-transduction pathway, T-DNA knockout mutants for AtCaM2, AtCaM3, and AtCaM4 were obtained and their thermotolerance tested. Of the three knockout mutant plants, there were no differences compared with wild type plants under normal conditions. However, only AtCaM3 knockout mutant showed a clear reduction in thermotolerance after heat treatment at 45°C for 50 min. Over-expression of AtCaM3 in either the AtCaM3 knockout or wild type background significantly rescued or increased the thermotolerance, respectively. Results from electrophoretic mobility-shift assays (EMSA), real-time quantitative RT-PCR, and western blot analyses revealed that, after HS, the DNA-binding activity of heat shock transcription factors (HSFs), mRNA transcription of heat shock protein (HSP) genes and accumulation of HSP were down-regulated in the AtCaM3 knockout mutant and up-regulated in AtCaM3-over-expressing transgenic lines. Taken together, these results suggest that endogenous AtCaM3 is a key component in the Ca2+-CaM HS signal-transduction pathway.




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