Worm Breeder's Gazette 12(5): 74 (February 1, 1993)

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Mapping and Sequencing of the Ryanodine Receptor Gene ryr-1 in Caenorhabditis Elegans

Yasuji Sakube, Hiroaki Kagawa

Dept. Biol., Fac. Sci., Okayama Univ. C51918 @JPNKUDPC.BITNET
Phone; 81-86-252-7865(new), Fax; 81-86-252-6601

To understand signal transduction mechanisms of excitation-contraction coupling, we cloned a cDNA of C. elegans ryanodine receptor gene by screening a cDNA library with a cDNA fragment of the rabbit skeletal ryanodine receptor as a probe(WBG 12#3, p51 ).The deduced 526 amino acids from 1.9 kb of cloned cDNA was ³47% identical to the C-terminus putative transmembrane segments M5 ~M10of mammalian ryanodine receptors. So we have concluded that the cloned cDNA encoded the calcium release channel region of the C. elegans ryanodine receptor. Southern blot analysis indicated that the gene was a single-copy. The cDNA mapped on the YAC Y44 Aand Y37G21 of chromosome V. The place was the same as that of cml6 c2 which was obtained by total cDNA project(Alan Coulson). Finally ryr-1 located in the cosmid clone of M04C11 .

Sequencing of recloned fragments from the cosmid M04C11 is progress and the result of up to date follows. cDNA sequences of our clones and cm16 c2 were found in the cosmid. Northern blot analysis shows that similar size(-15kb) as mammals of the transcript was detected. Deduced amino acid sequence of foot region had also high homology as was found transmembrane region. This conserved amino acid sequence of ryanodine receptor molecule suggest that foot structure might also have important function in signal transduction of excitation-contraction coupling. We are not known whether the complete gene of ryr-1 was in the single cosmid or expand to uncloned gap region although there were small introns in this gene as was common in C elegans. Ed Maryon and Phil Anderson are searching possibility of some corresponding mutants in that region(WBG 12#4, personal communication). We are makeing antibody against transmembrane region of the molecule by processing expressed protein from cloned cDNA in E. coli.