Molecular characterization of a 70 kDa heat-shock protein of bean mitochondria.
A bean cDNA clone that specifies a 70 kDa heat-shock protein (hsp70) has been isolated and sequenced. The nucleotide sequence analysis shows that the cDNA could encode a 72 kDa protein that is highly related to prokaryotic and mitochondrial members of the hsp70 family. The predicted protein was found to contain an amino-terminal extension typical of transit sequences. The in vitro transcription/translation product of the cDNA behaved as a 72 kDa polypeptide as predicted from the longest open reading frame. This polypeptide could be imported into isolated mitochondria and recovered as a 68 kDa product. The imported protein is identical in size to a mitochondrial protein that cross-reacts with hsp70-specific antibodies. The import data and Western blot analysis suggest that the cDNA clone encodes a mitochondrial member of the hsp70 family. Electrophoretic and immunoblot analysis reveal that the protein is loosely associated to the mitochondrial envelope and also exists as discrete soluble protein aggregates of about 270 and 420 kDa. Hsp70 of bean mitochondria can be in vitro phosphorylated on threonine residues in a calcium-dependent manner, and the modified protein was detected as an oligomer of about 160 kDa only. The data are discussed with respect to the chaperone function of hsp70 in mitochondria.
Vidal
V
V
Centre de Physiologie Végétale, URA CNRS no. 1457, Université Paul Sabatier, Toulouse, France.
Ranty
B
B
Dillenschneider
M
M
Charpenteau
M
M
Ranjeva
R
R
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D004247:DNA; D007887:Fabaceae / Q000235:genetics*; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D010946:Plants, Medicinal; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism
D006360:Heat-Shock Proteins; D010940:Plant Proteins; D011498:Protein Precursors; D004247:DNA