Worm Breeder's Gazette 16(1): 53 (October 1, 1999)

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.

Analysis of candidates for the factors that interact with UNC-51

Kenji Kawano1,2, Ken-ichi Ogura3, Yasumi Ohshima1

1 Department of Biology, Graduate School of Sciences, Kyushu University, Fukuoka 812-8581, Japan
2 kkawascb@mbox.nc.kyushu-u.ac.jp
3 CREST, JST, and Genome Biology Lab., National Institute of Genetics, Mishima 411-8540, Japan

     unc-51 and unc-14 mutants show abonormal axonal elongation and structures. Previously, we reported that the unc-51 gene encodes a novel Ser/Thr kinase and that its expression is observed in many neurons at all stages1). To analyze the function of UNC-51 further, we searched for proteins interacting with UNC-51 in the yeast two-hybrid system and found 16 clones including those for UNC-51 itself(one clone) and UNC-14(four clones). Identification of the unc-14 gene and direct interaction between UNC-51 and UNC-14 were also reported2). We are interested in the three clones, THB7, THB28 and THB47 among the remaining 11. Here we show the present status of the analysis .
     THB7 encodes a 2.8 kb transcript with SL1. This transcript encodes a 772 amino acid protein, which has 46% amino acid identity to human mitogen- and stress-activated protein kinase-1(MSK-1). MSK-1 contains two protein kinase domains in a single polypeptide and is activated by MAPK2/ERK2 or SAPK2/p38 in vitro3). The genomic gene for THB7 is located in cosmid C54G4(LG I).
     THB28 encodes a 1.4 kb taranscript with SL2. This transcript encodes a 316 amino acid protein, which has 46% amino acid identity to human EB1. EB1 was identified as a protein that binds to the product of a tumor suppressor gene APC(adenomatous polyosis coli)4). Another cDNA clone for THB28(ML12) was previously identified in our laboratory in a screen for proteins interacting with UNC-14(Shirakawa et al., unpublished). Therefore, this EB1 homologue THB28 could possibly interact with both UNC-51 and UNC-14. Many other cDNA clones such as yk417e3 and yk492c7 corresponding to THB28 have been obtained by Y. Kohara. The THB28 sequences are found in regions of Y59A8 YAC clone(LG I) that are apparently separated by more than 200kb.
     THB47 likely encodes a protein, which has approximately 40% amino acid identity to stress activated kinase JNK(c-Jun N-terminal kinase). The exact 5' end of THB47 cDNA is not yet determined. The genomic gene is found on cosmid T07A9(LG IV).
     We are constructing gfp fusion proteins and examining their expression patterns. Preliminary data suggest that THB7 is expressed in neurons, consistent with the result that unc-51 is expressed in many neurons. We also plan to confirm interaction of UNC-51 with THB7, 28 and 47 in an in vitro binding assay.

1) K. Ogura, C. Wicky, L. Magnenat, H. Tobler, I. Mori, F. M Müller, and Y. Ohshima : Genes & Dev., 8, 2389-2400, 1994
2) K. Ogura, M. Shirakawa, T. M. Barnes, S. Hekimi, and Y. Ohshima : Genes & Dev., 11, 1801-1811, 1997
3) M. Deak, A. D. Clifton, J. M. Lucocq, and D. R. Alessi : EMBO J., 17, 4426-4441, 1998
4) L.-K. Su et al. : Cancer Res., 55, 2972-2977, 1995