We have cloned a cDNA encoding a DnaJ-like protein from the human fibrosarcoma HT-1080 cDNA library. This cDNA encodes a protein of 397 amino acid residues whose sequence shows 38.2% identity with the Escherichia coli DnaJ protein and 47.2% with the yeast DnaJ homologue along the entire length. Since the sequence contains the N-terminal domain region conserved in DnaJ family members and the four repeats of the Cys-X-X-Cys-X-Gly-X-Gly motif which are characteristic of DnaJ proteins, we conclude that this cDNA clone encodes the human homologue of DnaJ.