Worm Breeder's Gazette 13(3): 98 (June 1, 1994)

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.

K252 acompound induced multiple anchor cells in N2 Hermaphrodites

Mitsunobu Hara[1], Min Han[2]

[1]Present address: Tokyo Research Lab. Kyowa Hakko, Machida, Tokyo 194, JAPAN
[2]Dept. of MCD Biology, University of Colorado at Boulder, CO 80309

K252 awas originally discovered as a potent inhibitor of cyclic nucleotide - dependent protein kinases and protein kinase C (Kase et al. B.B.R.C. 142, 436, 1987). It has recently been reported that K252 ais a potent and selective inhibitor of NGF receptor / trk kinase (Tapley et al. Oncogene 7, 371, 1992). We have examined the effect of K252 aon N2 hermaphrodites, and found that it induces sterile and uncoordinated phenotypes at 18 to 35 nmol of drug / plate. In order to gain further insight into the drug-induced phenotype, we studied K252 a-treatedworms under Nomarski optics and found that K252 ainduced two or three anchor cells in N2 worms in a dose dependent manner, suggesting that K252 aaffects the VU-AC decision of Zl.ppp and Z4 .aaacells in such a way that those cells both adopt the AC fate. K252 ahas no effect on lin-12 (d, n137 ). That is, lin-12 ( n137 )has no anchor cell and has multiple pseudovulvae even in the presence of 35nmol of K252 a.It is therefore likely that the inhibitory target of K252 amight be upstream of lin-12 . let-60 (gf, n1046 )is resistant to K252 a. K252 a(35nmol)-treated let-60 ( nlO46 )is fertile and is not uncoordinated. Furthermore, 95% of let-60 ( n1046 )have 1 anchor cell in the presence of high concentrations of K252 a.In mammalian neural cells such as rat pheochromocytoma PC12 cells, ras functions downstream of the NGF-receptor. Supposing that the inhibitory target of K252 awould be an NGF-receptor in C. elegans, the above result would imply that let-60 could function downstream of the NGF receptor of C. elegans. Toward understanding the molecular target of K252 ain C. elegans, we screened for K252 a-resistantstrains, and isolated one resistant mutant, ku104 . ku104 is completely resistant to 35 nmol of K252 a.Genetic mapping of ku104 is in progress. We are also trying to isolate K252 a-sensitivemutants with the aim of isolating K252 a-sensitivealleles of the C. elegans NGF-receptor