Early evolution of metazoan serine/threonine and tyrosine kinases: identification of selected kinases in marine sponges

Abstract

The phylum Porifera (sponges) was the first to diverge from the common ancestor of the Metazoa. In this study, six cDNAs coding for protein-serine/threonine kinases (PS/TKs) are presented; they have been isolated from libraries obtained from the demosponges Geodia cydonium and Suberites domuncula and from the calcareous sponge Sycon raphanus. Sequence alignments of the catalytic domains revealed that two major families of PS/TK, the "conventional" (Ca(2+)-dependent) protein kinase C (PKC), the cPKC subfamily, as well as the "novel" (Ca(2+)-independent) PKC (nPKC), form two separate clusters. In each cluster, the sequence from S. raphanus diverges first. To approach the question about the origin of protein-tyrosine kinases (PTK), which are found only in Metazoa, we analyzed two additional PS/TKs which have been cloned from S. domuncula: the stress-responsive protein kinase (KRSvSD) and the protein-kinase-C-related kinase (PRKvSD). The construction of the phylogenetic tree, comprising the eight PS/TKs and the PTK cloned previously from G. cydonium, revealed that the PTK derived from the branch including the KRSvSD kinase. These data facilitate the first molecular approach to elucidate the origin of metazoan PTK within the PS/TK superfamily.

Formation of giant spicules in the deep-sea hexactinellid Monorhaphis chuni (Schulze 1904): electron-microscopic and biochemical studies

“…Recent molecular biological studies have revealed that the siliceous demosponges are provided with the genetic "toolkit" to construct these skeletal elements, the spicules, functional animals being composed of differentiated cells that interact in a tissue-like manner and that allow the formation of a body plan (Müller et al 2004;Müller 2005;Pilcher 2005). M. chuni is a siliceous sponge, belonging to the phylogenetically oldest Poriferan class, the Hexactinellida (Kruse et al 1997). …”

“…Based on molecular biological (Kruse et al 1997) and geological (Reitner andestablished as the oldest, having evolved between Neoproterozoic glaciations (720-585 Ma), sometimes referred to as "snowball" Earth (for a review, see Corsetti et al 2006). During this period, a cycle of silicate weathering and carbonate precipitation, prior to or simultaneously with the glaciations, has been proposed to have resulted in the dissolution of surface rocks composed of insoluble silicates (CaSiO 3 ).…”

Sponge Bcl-2 homologous protein (BHP2-GC) confers distinct stress resistance to human HEK-293 cells

“…50,51 In addition, FuGENE 6 reagent, PCR-DIG (digoxigenin)-ProbeSynthesis Kit, anti-DIG AP Fab fragments and CDP [disodium 2-chloro-5- (4-methoxyspiro{1,2-dioxetane-3,2'-(5'-chloro)-tricyclo [3.3.1.1. 3,7 ]decan}-4-yl)phenyl phosphate] from Roche Diagnostics (Mannheim; Germany); RPMI-1640 and OptiMEM from Life Technologies (Karlsruhe; Germany); geneticine G418 from Invitrogen (Groningen; The Netherlands); tributyltin and labelled anti-rabbit Ig from Sigma (Deisenhofen; Germany); AC-DEVD-AMC from Peptide Institute (Osaka; Japan); monoclonal anti-Bcl-2 [human] antibodies (mouse) from Santa Cruz (Santa Cruz, Cal; USA); anti-mouse (Cy3 labeled) and anti-rabbit (FITC-labeled) from Dianova (Hamburg; Germany); the mouse anti-Hexa-histidine monoclonal antibody (McAb-anti-His) from Invitrogen (Groningen; The Netherlands).…”

Identification of a silicatein(-related) protease in the giant spicules of the deep-sea hexactinellidMonorhaphis chuni

“…For the isolation of the cDNA encoding cystatin, a cDNA library of Suberites domuncula was used (Kruse et al, 1997). From this a set of 30·000 expressed sequence tag sequences has been compiled.…”