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In-vivo and in-vitro synthesis of photosynthetic fructose-1,6-bisphosphatase from pea (Pisum sativum L.)

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Abstract

Etiolated pea (Pisum sativum L. cv. Lincoln) seedlings do not show any capability for the biosynthesis of chloroplast fructose-1,6-bisphosphatase (FBPase), but the rate of biosynthesis of the increases with the pre-illumination time. This light-induced FBPase synthesis appears to be regulated at the transcriptional level, the response of young leaves being greater than that of mature ones. In-vivo labelling experiments demonstrated by immunoprecipitation, followed by sodium dodecyl sulfate electrophoresis and fluorography, the presence of a 49-kilodalton (kDa) band which corresponds to the mature FBPase subunit. In-vitro translation experiments with a wheat-germ synthesizing system and polyadenylated mRNA isolated from illuminated young pea seedlings have demonstrated the appearance of a 59-kDa labelled band corresponding to the precursor of the FBPase basic subunit. When intact pea chloroplasts were added to the above in-vitro incubation mixture, a labelled 49-kDa subunit similar to that of the in-vivo experiments appeared in the organelle under illumination. From these results we can conclude that a 10-kDa transit peptide bound to the translated pea FBPase subunit exists in the cytosol; this transit peptide is lost during passage through the chloroplast envelope, leaving the mature subunit inside the organelle.

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Abbreviations

ELISA:

enzyme-linked immunosorbent assay

FBPase:

fructose-1,6-bisphosphatase

kDa:

kilodalton

MW:

molecular weight

PBS:

phosphate-buffered saline

PMSF:

phenylmethylsulfonyl fluoride

poly(A)mRNA:

polyadenylated mRNA

SDS:

sodium dodecyl sulfate

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Authors and Affiliations

  1. Department of Plant Biochemistry, Estación Experimental del Zaidín (CSIC), Profesor Albareda 1, E-18008, Granada, Spain

    M. Sahrawy, A. Chueca, R. Hermoso, J. J. Lázaro & J. López Gorgé

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  1. M. Sahrawy
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  2. A. Chueca
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  3. R. Hermoso
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  4. J. J. Lázaro
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  5. J. López Gorgé
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Sahrawy, M., Chueca, A., Hermoso, R. et al. In-vivo and in-vitro synthesis of photosynthetic fructose-1,6-bisphosphatase from pea (Pisum sativum L.). Planta 182, 319–324 (1990). https://doi.org/10.1007/BF02411381

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  • DOI: https://doi.org/10.1007/BF02411381

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