Dictyostelium discoideum contains a family of calmodulin-related EF-hand proteins that are developmentally regulated

Abstract

A full-length genomic DNA fragment that codes for a novel EF-hand protein from Dictyostelium discoideum was cloned and sequenced. The protein is composed of 168 amino acids and contains four consensus sequences that are typical for Ca 2+-binding EF-hand domains. The protein sequence exhibits only minor similarities to other calmodulin-type proteins from Dictyostelium. The genomic DNA harbors two short introns; their positions suggest that the gene is unrelated to the EF-hand proteins from the calmodulin group. Northern blot analysis showed that the mRNA level was significantly increased during development. Polyclonal antibodies raised against the recombinant protein recognized in Western blots a protein of about 20 kDa. Like the mRNA, also the protein was more abundant in developing cells. Overlay experiments with 45Ca 2+ indicated that the EF-hands in fact have Ca 2+-binding activity. The recent description of CBP1, another calmodulin-type Dictyostelium protein that is upregulated during development [Coukell et al. (1995) FEBS Lett. 362, 342–346], suggests that D. discoideum contains a family of EF-hand proteins that have specific functions during distinct steps of development. We therefore designate the protein described in this report as CBP2.