1. Groucho suppresses Pax2 transactivation by inhibition of JNK-mediated phosphorylation The EMBO Journal Vol. 22 No. 20 pp. 5522-5529, 2003

2. Groucho -wnt/b-cat

3. -human Groucho family: TLE-1/2/3/4 (transducin-like enhancer of split) hAES (amino-termianl enhancer of split) -mouse Groucho family: Grg-1/2/3/4 & Grg-5 Q domain: glutamine rich GP domain: glycine/proline rich CcN domain SP domain:serine/proline rich Four WD40 repeats domain tetramerization (leucine zipper-like structure) binding to n-terminal of H3 Interaction with HDAC Putative casein kinase II/ cdc2 phosphorylation sites nuclear localization signal protein interaction

4. Pax: paired box gene -pax 1~9, DNA binding transcription factor -activation and repression of downstream target genes -regulators of organogenesis & key factors in maintaining pluripotency of stem cell populations during development -Mutations of the Pax: profound developmental defects in organisms as diverse as flies, mice and humans.

5. PD domain: bipartite paired domain N- & C-terminal subdomains three α-helices third helix of each subdomain (red) contact with the major groove of DNA OP motif: octapeptide motif HD domain: homeodomain helix–turn–helix structure. TD domain: C-term transactivation domain

6. Pax2/5/8 -the Pax2/5/8 proteins contain a conserved octapeptide with homology to the engrailed-type repressor sequence. OP domain Engrailed homology region of the homeodomain transcription factors EN2 and Goosecoid(GSC) -pax 2/5/8 contains partial homeodomain(HD1) that consistutes an interation surface for both the retinoblastoma(Rb) gene product and the TATA-binding protein (Eberhard and busslinger,1999)

7. - The ability of Pax2 to activate transcription is enhanced by phosphorylation of its activation domain by the JNK (Cai et al., 2002) Pax2-JNK-groucho - Pax5 transactivation activity can also be negatively regulated through interactions with the Groucho/TLE family of co-repressors (Eberhard et al.,2000) -In mouse and man, the Pax2 gene is essential for the development of the kidneys, optic cup, and inner ear. -SFRP2 gene is a target of the Pax2 transcription factor (J Biol Chem.2003 )

8. Fig1. Grg4 physically interacts with Pax2

9. TSA(trichostatin A): HDAC inhibitor Fig. 2. Grg4 inhibits the JNK enhanced transactivation potential of Pax2. Gal4 DNA binding element luci Gal 4 DNA binding domain Activation domain Elk or c-Jun

10. Fig. 3. Grg4 inhibits phosphorylation of Pax2 in a DNA-dependent manner In vivo phospho-labeling of 293 cells

11. Fig. 4. Co-expression of Pax2 and Grg4 in the embryonic kidney and neural tube. E-cadherin (green) anti-Pax2 (red) anti-Grg4 (red) E15.5 embryos. immunostaining Kidney Spinal cord s-shaped body (arrow). renal vesicles (arrowhead) embryonic pancreas E16 tissue/ NE