Characterization of a novel drought-induced 34-kDa protein located in the thylakoids of Solanum tuberosum L. plants
1996
Pruvot, G. | Cuine, S. | Peltier, G. | Rey, P.
Using two-dimensional electrophoresis and Coomassie Blue staining, the accumulation of a 34-kDa protein (named cdsp 34 for chloroplastic drought-induced stress protein) is shown in the thylakoids of Solanum tuberosum plants subjected to a progressive and reversible water deficit. In-vivo labeling experiments showed an increased synthesis of cdsp 34 from the early stages of drought stress (leaf relative water content around 85%) and throughout the constraint. Sequences of the N-terminal part and of four tryptic-digest peptides did not reveal significant homology between the cdsp 34 protein and other known proteins. Western blotting analysis, using a serum raised against the N-terminal part of cdsp 34, confirmed the accumulation of cdsp 34 in thylakoids upon drought stress. From immunoblot analysis of different chloroplastic subfractions, the cdsp 34 protein appears to be an extrinsic protein preferentially located in unstacked stroma thylakoids. Immunoprecipitation of in-vitro-translated products, as well as Southern analysis, showed that the cdsp 34 protein is nuclear encoded. After rewatering of water-stressed plants, the level of cdsp 34 synthesis was reduced, but remained substantially higher than in control plants. Western analysis showed the persistence of a high amount of cdsp 34 in rewatered plants for at least two weeks. Based on the abundance and on the location of cdsp 34 within thylakoids, a putative role for this novel chloroplastic protein is discussed in relation to the tolerance of the photosynthetic apparatus of higher plants to dehydration.
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