Molecular evolution of apoptotic pathways: Cloning of key domains from sponges (Bcl-2 homology domains and death domains) and their phylogenetic relationships

Cells from metazoan organisms are eliminated in a variety of physiological and pathophysiological processes by apoptosis. In this report, we describe the cloning and characterization of molecules from the marine sponges Geodi a cydonium and Suberites domuncula, whose domains show a high similarity to those that are found in molecules of the vertebrate Bcl-2 superfamily and of the death receptors. The Bcl-2 proteins contain up to four Bcl-2 homolog y regions (BH). Two Bcl-2-related molecules have been identified from spong es that are provided with two of those regions, BH1 and BH2, and are termed Bcl-2 homology proteins (BHP). The G. cydonium molecule, BHP1_GC, has a pu tative size of 18,164, while the related sequence from S. domuncula, BHP1_S D, has a M-r of 24,187. Phylogenetic analyses of the entire two sponge BHPs revealed a high similarity to members of the mammalian Bcl-2 superfamilies and to the Caenorhabditis elegans Ced-9. When the two domains. BH1 and BH2 , are analyzed separately, again the highest similarity was found to the me mbers of the Bcl-2 superfamily, but a clearly lower relationship to the C. elegans BH1 and BH2 domains in Ced-9. In unrooted phylogenetic trees the sp onge BH1 and BH2 are grouped among the mammalian sequences and are only dis tantly related to the C. elegans BH domains. The analysis of the gene struc ture of the G. cydonium BHP showed that the single intron present is locate d within the BH2 domain at the same position as in C. elegans and rat Bcl-x (L). In addition, a sponge molecule comprising two death domains has been c haracterized from G. cydonium. The two death domains of the potential proap optotic molecule GC_DD2, M-r 24,970, share a high similarity with the Fas-F ADD/MORT1 domains. A death domain-containing molecule has not been identifi ed in the C. elegans genome. The phylogenetic analysis revealed that the sp onge domain originated from an ankyrin building block from which the mammal ian Fas-FADD/MORT1 evolved. It is suggested that the apoptotic pathways tha t involve members of the Bcl-2 superfamily and of the death receptors are a lready present in the lowest metazoan phylum, the Porifera.