Molecular cloning and characterization of ryanodine receptor from unfertilized sea urchin eggs.
Unfertilized eggs of sea urchins (Hemicentrotus pulcherrimus) demonstrated cyclic ADP-ribose (cADPR)-induced Ca(2+) release and caffeine-induced Ca(2+) release, both of which were considered to be mediated through the ryanodine receptor (RyR). We cloned cDNAs for sea urchin egg RyR (suRyR), which encode a 597-kDa protein of 5,317 amino acids. suRyR shares common structural features with known RyRs: the well-conserved COOH-terminal domain, which forms a functional Ca(2+) channel, and a large hydrophilic NH2-terminal domain. suRyR shows amino acid sequence identity (43-45%) similar to the three mammalian RyR isoforms. Phylogenetic analysis indicates that suRyR branched from three isoforms of vertebrates before they diverged, suggesting that suRyR may be the only RyR isoform in the sea urchin. Four in-frame insertions were found in suRyR cDNAs, one of which was novel and unique, in that it had a cluster of serine residues. The transcripts with and without these insertions were found in the egg RNA. These results suggest that suRyR may be expressed as a functional Ca(2+)-induced Ca(2+) release channel, which might also be involved in cADPR-induced Ca(2+) release.
Shiwa
Mieko
M
Department of Pharmacology, Juntendo University School of Medicine, Tokyo 113-8421, Japan.
Murayama
Takashi
T
Ogawa
Yasuo
Y
D000246:Adenosine Diphosphate Ribose / Q000031:analogs & derivatives* / Q000494:pharmacology; D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D036563:Cyclic ADP-Ribose; D018076:DNA, Complementary / Q000235:genetics; D005306:Fertilization; D008969:Molecular Sequence Data; D005810:Multigene Family; D010063:Ovum / Q000378:metabolism*; D010802:Phylogeny; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics* / Q000378:metabolism*; D012617:Sea Urchins / Q000378:metabolism*
D018076:DNA, Complementary; D019837:Ryanodine Receptor Calcium Release Channel; D036563:Cyclic ADP-Ribose; D000246:Adenosine Diphosphate Ribose; D002110:Caffeine; D002118:Calcium