Then, the activated CDPK would regulate the activation of NADPH oxidase in the plasma membrane and release ROS. Finally, downstream of ROS production, the UV B induced and activated MAP kinases possibly participate http://www.selleckchem.com/products/CAL-101.html in the activation of regulatory proteins such as GT 1 nuclear factor leading to transcriptional activation of TIA biosynthetic genes and enhanced production of catharan thine. It has been earlier reported that yeast elicitor in C. roseus activates the octadecanoid pathway. leading to an increase in jasmonic acid levels via the activation of calcium influx and protein phosphorylation cascades. JA induces the expression of the ORCA3 gene via post translational modification which further interacts with the Tdc promoter and the YE and JA responsive RV frag ment of the Str promoter enhancing the gene expression.

YE reportedly also induce the expression of the zinc finger proteins, which by binding to specific elements within the promoter regions of Tdc and Str can repress its gene expression. Similarly YE induced CrBPF1 expression has been reported to be putatively involved in the regulation of STR via interaction with the BA region. It would be interesting to understand whether the UV B and YE induced TIA pathway share common ele ments in signal transduction and also if UV B utilizes any of the transcriptional initiators or repressors induced by YE in initiating the TIA pathway. Methods Chemicals 2, 7 DCFH DA, EGTA, heparin, histone IIIS, N acetyl cysteine, phosphothreonine, phosphotyrosine, sodium fluoride, sodium orthovanadate and verapamil were pur chased from Sigma Chemical Company, St.

Louis, USA. Sodium glycerophosphate and sodium fluoride were from Hi media Laboratories, India. Catharanthine and vindoline were obtained from Shanghai kangai biologi cals, China. Staurosporine and suramin were obtained from MP Biomedicals, Germany. Monoclonal antibodies to phospho serine and phospho tyrosine, complete pro tease inhibitor cocktail and myelin basic protein were pur chased from Upstate laboratories, U. S. A. SB 203580, PD 98059 and SB 600125 were a kind gift from Prof. Anjali Karande, I. I. Sc, Bangalore. Cell culture and treatments of cells with UV B and chemicals C. roseus suspension cultured cells were cultivated as described previously. A three ml of six day old cul ture in stationary growth phase was transferred aseptically to 35 mm petri plates and irradiated with UV B directly, at a dis tance of 2.

5 cm between the cultured cells and the lamp as described. For chemical treatments, agonists or antagonists of effectors involved in other signal transduc tion pathways were diluted in water to the appropriate final concentrations, as indicated in figure legends from stock solutions prepared as described Anacetrapib in Table 1. The cells were treated for 10 min with different chemicals and subsequently irradiated with UV B for 5 min, as indicated in figure legends.