Using a multistep system for protein purification, including affinity chromatography on zeatin riboside–Sepharose, we isolated a zeatin-binding protein from the chloroplasts of the first leaf of ten-day-old barley seedlings (ZBPChl). In combination with trans-zeatin, this protein enhanced the RNA synthesis in the chloroplast lysate from the same leaves. A hormonal specificity of ZBPChl-induced transcriptional stimulation was demonstrated. The protein activated the chloroplast but not nuclear genome. Age-dependent changes in the ZBPChl functional activity and in the response of the chloroplast transcription system to this protein and trans-zeatin were observed. In the chloroplast lysate from aging first leaves of 14-day-old barley plants, ZBPChl from the leaves of 3-, 10-, and 14-day-old barley plants activated transcription in the absence of the hormone, and trans-zeatin suppressed this activation. Thus, trans-zeatin played the role of the ZBPChl activator in chloroplast lysates from ten-day-old seedling and its inhibitor in lysates from 14-day-old seedlings. The combination of ZBPChl and trans-zeatin activated the RNA synthesis in lysates from lupine leaves, which indicates the absence of species-specificity in the ZBPChl action. It was shown that, in the heterologous transcription system, the principal factor of regulation was not a plant species, which was a source of the ZBPChl or chloroplast lysate, but the physiological state of plants used.
Russian Journal of Plant Physiology – Springer Journals
Published: Oct 13, 2004
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