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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Calmodulin Is Involved in Heat Shock Signal Transduction in Wheat¹

Institute of Molecular Cell Biology, Hebei Normal University, Shijiazhuang 050016, People's Republic of China (H.-T.L., B.L., Z.-L.S., R.-L.M., D.-Y.S., R.-G.Z.); and Institute of Agro-physics, Physiology, and Biochemistry, Hebei Academy of Agricultural Sciences, Shijiazhuang 050051, People's Republic of China (H.-T.L., B.L., X.-Z.L., R.-L.M., R.-G.Z.)

The involvement of calcium and calcium-activated calmodulin (Ca²⁺-CaM) in heat shock (HS) signal transduction in wheat (Triticum aestivum) was investigated. Using Fluo-3/acetoxymethyl esters and laser scanning confocal microscopy, it was found that the increase of intracellular free calcium ion concentration started within 1 min after a 37°C HS. The levels of CaM mRNA and protein increased during HS at 37°C in the presence of Ca²⁺. The expression of hsp26 and hsp70 genes was up-regulated by the addition of CaCl₂ and down-regulated by the calcium ion chelator EGTA, the calcium ion channel blockers LaCl₃ and verapamil, or the CaM antagonists N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide and chlorpromazine. Treatment with Ca²⁺ also increased, and with EGTA, verapamil, chlorpromazine, or trifluoperazine decreased, synthesis of HS proteins. The temporal expression of the CaM1-2 gene and the hsp26 and hsp70 genes demonstrated that up-regulation of the CaM1-2 gene occurred at 10 min after HS at 37°C, whereas that of hsp26 and hsp70 appeared at 20 min after HS. A 5-min HS induced expression of hsp26 after a period of recovery at 22°C after HS at 37°C. Taken together, these results indicate that Ca²⁺-CaM is directly involved in the HS signal transduction pathway. A working hypothesis about the relationship between upstream and downstream of HS signal transduction is presented.

¹ This work was supported by the National Natural Science Foundation of China (grant no. 3977075), by the Natural Science Foundation of Hebei Province, China (grant no. 301447), and by the National Key Basic Research Special Funds, China (G1999011700).

^* Corresponding author; e-mail zhourengang{at}163.com; fax 0086-311-7042490.

Received December 6, 2002; returned for revision January 14, 2003; accepted March 17, 2003.

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