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hindi songs download hd 720pBlocking of (3′-phosphoryl) AMP and poly(I)-poly(C) stimulation of chloroplast RNA synthesis in isolated pea chloroplasts.
Chloroplasts isolated from pea (Pisum sativum L.) were permeabilized by detergent treatment to trap nucleotide triphosphates and inorganic pyrophosphate in the internal space. Chloroplasts incubated with phosphoryl analogues of nucleoside triphosphates (ATP.CTP and ATP.GTP) and the ribonucleotide (1′-RpATP).CTP and 1′-RpATP.GTP (Rp = (3′-phosphoryl) adenosine) generated in the chloroplast stroma on addition of Mg-ATP as substrate and [32P]GTP by endogenous CTP: 1′-RpATP.CTP ribotransferase (EC 2.7.7.25). No 32P label was observed in the chloroplast stroma on the addition of [32P]CTP, ATP, ATP.CTP, or ATP.GTP alone, indicating a requirement for addition of CTP to generate a CTP product that can be formed via the ribotransferase reaction. The generation of 32P label in the stroma from [32P]CTP in the presence of ATP or ATP.CTP was half-maximal at a level of about 0.8 microM of the appropriate nucleotide analogue, and in the presence of poly(I)-poly(C) stimulation of RNA synthesis was half-maximal at about 6 microM of analogue. A Mg-ATP concentration of about 2 mM proved optimal for this stimulation, and the level of stimulation was of the same order of magnitude as found with chloroplasts permeabilized with anionic detergents. ATP analogues with no alpha, beta-gamma phosphoryl groups (AMP.CTP and AMP.GTP) were unable to support stromal RNA synthesis. In contrast, pyrimidine and purine nucleotides were found to support similar levels of stromal RNA synthesis as ATP. The Mg-ATP requirement and the effects of nucleotides with alpha and beta phosphoryl groups on the level of stromal RNA synthesis were similar to those previously demonstrated for

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