Characterisation of PHSP1, a cDNA encoding a mitochondrial HSP70 fromPisum sativum

Abstract

A pea cDNA clone,PHSP1, encoding a member of the HSP70 gene family has been isolated. DNA sequence analysis indicates that the protein encoded byPHSP1 is a homologue of the mitochondrial HSP70 proteins, SSP1 fromSchizosaccharomyces pombe and SSC1 fromS. cerevisiae. It contains an amino-terminal extension of 50 amino acids, rich in basic and hydroxyl amino acids, similar to other plant mitochondrial leader sequences. Western blot analysis indicates that the PHSP1 protein is associated only with mitochondria and not with any other sub-cellular organelle or cytoplasm. Further confirmation of its location within mitochondria was obtained fromin vitro protein translocation experiments into purifiedPisum sativum mitochondria. It was observed that the precursor protein was efficiently imported and that it is processed to produce a protein with anM_r of the anticipated size of the mature protein. Results are discussed with respect to the structure and function of the mitochondrial HSP70 protein.