Characterization of Chloroplast Clp proteins in Arabidopsis: Localization, tissue specificity and stress responses.
The ATP-dependent Clp protease is one of the newly identified proteolytic systems in plant organelles that incorporate the activity of molecular chaperones to target specific polypeptide substrates and avoid inadvertent degradation of others. We describe new nuclear-encoded ClpC (ClpC1) and ClpP (ClpP3-5) isomers in Arabidopsis thaliana that raise the total number of identified Clp proteins to 19. The extra Clp proteins are localized within the stroma of chloroplasts along with the ClpD, -P1 and -P6 proteins. Potential differential regulation among these Clp proteins was analysed at both the mRNA and protein level. A comparison between different tissues showed increasing amounts of all plastid Clp proteins from roots to stems to leaves suggested the greatest abundance of proteins was in chloroplasts. The increases in protein were mirrored at the mRNA level for most ClpP isomers (ClpP1, -3, -4 and -6) but not for the three Hsp100 proteins (ClpC1, -C2 and -D) and ClpP5, which exhibited little change in transcript levels, suggesting post-transcriptional/translational regulation. Potential stress induction was also tested for all chloroplast Clp proteins by a series of brief and prolonged stress conditions. Short-term moderate and severe stresses (desiccation, high salt, cold, heat, oxidation, wounding and high light) all failed to elicit significant or rapid increases in any chloroplast Clp protein. However, increases in mRNA and protein content for ClpD and several ClpP isomers did occur during long-term high light and cold acclimation of Arabidopsis plants. These results reveal the great complexity of Clp proteins within the stroma of plant chloroplasts, and that these proteins, rather than being rapidly induced stress proteins, are primarily constitutive proteins that may also be involved in plant acclimation to different physiological conditions.
Zheng
Bo
B
Umeå Plant Sciences Centre, Department of Plant Physiology, Umeå University, SE-901 87 Umeå, Sweden Department of Agricultural Botany, Faculty of Agriculture, Hebrew University of Jerusalem, Rehovot 76100, Israel Department of Biochemistry, Institute of Organic Chemistry and Biochemistry, Flemingovo nám.2, C2-16610 Prague 6, Czech Republic Present address: Botanical Institute, Göteborg University, Box 461, SE-405 30 Göteborg, Sweden.
Halperin
Tami
T
Hruskova-Heidingsfeldova
Olga
O
Adam
Zach
Z
Clarke
Adrian K
AK