Worm Breeder's Gazette 14(2): 79 (February 1, 1996)
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.
Lab. of Molecular Biology. Toyohashi University of Technology, Toyohashi 441, Japan.
We have previously reported the transformation rescue of the emb-8(hc69) mutant with a cosmid containing the gene cej-1(C. elegans Meeting 1995, abstract 310). emb-8 is an embryonic lethal mutant at nonpermissive temperature (25oC). Cell division arrest between one cell to 50 cells stage. The egg shell is osmotically sensitive. Recently we have come to know from Sarah Crittenden( Judith Kimble Lab.) that emb-8 mislocalizes Glp-1 & P-granules specific antigens. We have earlier observed symmetric distribution of P-granules in a fraction of embryos using a monoclonal antibody specific to germ cells. These observation suggest a role of emb-8 in early cell divisions of the embryos. We have narrowed down the rescueing length of the DNA. It is about 5.5 kb which contain about 2.5 kb coding sequence and arround 3 kb noncoding upstream sequence. The rescuing ability of the 5.5 kb genomic DNA fragment appears less than when the whole cosmid ZC235 is used in the transformation experiment.This 5.5 kb fragment contains a single gene which we named as cej-1. This gene encodes a novel protein of 600 a.a. which have no significant homology with any protein in the data bank. It shows about 30% identity with calphotin, spasmolysin precursor, Hypothetical Protein 2280, in a stretch of about 200 a.a. & about similar identity with Endochitinase, Glucan 1,4-alpha-Glucosidase, GVPC protein, Hydroxy proline-rich glycoprotein precursor, mRNA cap binding protein, Probable cell surface glycoprotein sed1, in a stretch of about 100 a.a. The CEJ-1 protein has three 70 a.a. repeats,six 30 a.a.repeats & three 20 a.a. repeats. Northern analysis suggest that the cej-1 expresses in L3 larvae, gradually increasing as the development proceeds reaching a maximum in adult animals. Signal is detected only in the early as we did not observe any signal after 1.5 hours of the first cleavage. We are further performing in situ and immunocytochemical studies on emb-8 embryos. We thank Sarah Crittenden for her unpublished results,and Glp-1 antibody; Kikuno for the analysis of the cej-1 protein structure; Yuji Kohara for cDNA clones; Alan Coulson for cosmid DNA; J. Miwa for advice and encouragement.