Regulated expression and phosphorylation of the 23-26-kDa ras protein in the sponge Geodia cydonium.
We have cloned, sequenced and examined the sponge Geodia cydonium cDNA encoding a protein homologous to ras proteins. The sponge ras protein has a more conserved N-terminal region and a less conserved C-terminal region, especially in comparison to Dictyostelium discoideum; the similarity to human c-Ha-ras-1 and to Saccharomyces cerevisiae is less pronounced. The sponge ras cDNA comprises five TAG triplets; at the translational level these UAG termination codons are suppressed by a Gln-tRNA. The sponge ras protein was isolated and partially purified (23-26 kDa) and found to undergo phosphorylation at a threonine moiety, when dissociated cells were incubated in the presence of a homologous aggregation factor and insulin. Insulin-mediated phosphorylation of the ras protein resulted in a decrease in its Kd with GTP from 2 microM to 80 nM. The activated ras protein displayed high GTPase activity if the partially purified protein was incubated with homologous lectin and lectin receptor molecules. These results suggest that in the sponge, ras is activated by the insulin/insulin(insulin-like)-receptor system. This transition enables the ras protein to interact with the lectin-receptor/lectin complex, a process which may ultimately lead to an initiation of an intracellular signal-transduction chain.
Abteilung Angewandte Molekularbiologie, Universität Mainz, Federal Republic of Germany.
A
A
Robitzki
H C
HC
Schröder
D
D
Ugarković
Y
Y
Kuchino
B
B
Kurelec
V
V
Gamulin
W E
WE
Müller
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation; D015723:Gene Library; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D011161:Porifera / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D007328:Insulin; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras)