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J. Biol. Chem., Vol. 280, Issue 23, 22335-22342, June 10, 2005

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Mind Bomb-2 Is an E3 Ligase for Notch Ligand^*

Bon-Kyoung Koo, Ki-Jun Yoon, Kyeong-Won Yoo, Hyoung-Soo Lim, Ran Song, Ju-Hoon So, Cheol-Hee Kim¶, and Young-Yun Kong||

From the Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang, Kyungbuk, 790-784 South Korea and the Department of Biology, Chungnam National University, Daejeon 305-764, South Korea

The zebrafish gene, mind bomb (mib), encodes a protein that positively regulates of the Delta-mediated Notch signaling. It interacts with the intracellular domain of Delta to promote its ubiquitination and endocytosis. In our search for the mouse homologue of zebrafish mind bomb, we cloned two homologues in the mouse genome: a mouse orthologue (mouse mib1) and a paralogue, named mind bomb-2 (mib2), which is evolutionarily conserved from Drosophila to human. Both Mib1 and Mib2 have an E3 ubiquitin ligase activity in their C-terminal RING domain and interact with Xenopus Delta (XD) via their N-terminal region. Mib2 is also able to ligate ubiquitin to XD and shift the membrane localization of Delta to intracellular vesicles. Importantly, Mib2 rescues both the neuronal and vascular defects in the zebrafish mib^ta52b mutants. In contrast to the functional similarities between Mib1 and Mib2, mib2 is highly expressed in adult tissues, but almost not at all in embryos, whereas mib1 is abundantly expressed in both embryos and adult tissues. These data suggest that Mib2 has functional similarities to Mib1, but might have distinct roles in Notch signaling as an E3 ubiquitin ligase.

Received for publication, February 11, 2005 , and in revised form, April 4, 2005.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY974090 and AY974091.

^* This work was supported by Vascular System Research Center Grant R11-2001-090-03001-0 from KOSEF, Basic Research Program of the Korea Science & Engineering Foundation Grant R02-2003-000-10057-0), the 21C Frontier Functional Human Genome Project from Ministry of Science and Technology of Korea Grant FG04-22-05), and Molecular and Cellular BioDiscovery Research Program Grant M1-0106-01-0001 from the Ministry of Science and Technology, South Korea. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains Fig. S1.

¶ To whom correspondence may be addressed. Fax: 82-42-822-9690; E-mail: zebrakim{at}cnu.ac.kr. || To whom correspondence may be addressed. Fax: 82-54-279-2199; E-mail: ykong{at}postech.ac.kr.

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