Molecular cloning of an intronless gene for the hamster centromere antigen CENP-B

Abstract

Centromere protein B (CENP-B) is a DNA-binding protein present at both active and inactive centromeres. It was first localized at the kinetochore region by human autoimmune sera from CREST patients. Using a previously identified human cDNA we have isolated a genomic clone containing the complete hamster CENP-b intronless coding sequence. At the nucleotide level it was found to possess a high degree of homology with the human and mouse CENP-B genes, being 75% and 90% respectively. This codes for 606 amino acid residues, which represent seven more than the human and mouse centromeric proteins. Hamster CENP-B protein analysis revealed at the N-terminal region a 133 amino acid fragment of 100% homology to the DNA binding motif identified previously for the human autoantigen. Expression of hamster CENP-B during the cell cycle was analyzed by using a specific antiCENP-B serum generated against the C-terminal conserved region. These data indicate that CENP-B is highly conserved and it represents a universal component of the centromere structure and function in mammals.