Tipin, a novel timeless-interacting protein, is developmentally co-expressed with Timeless and disrupts its self-association

The mouse Timeless gene (mTim) was identified originally on the basis of its similarity to a Drosophila circadian gene, but has no substantiated role in the circadian clock mechanism. The importance of mTim in cellular processes involved in development, however, is undeniable, since targeted mutagenesis of this gene arrests embryonic development. To connect mTim to known pathways controlling cellular processes important for early development, a yeast two-hybrid approach was used to identify embryonic mTIM-interacting proteins. One positive interactor, a previously uncharacterized protein that is here termed TIPIN (TIMELESS interacting protein), was shown to interact with mTIM in vitro and in cultured cells. mTim and Tipin transcripts are co-expressed in similar tissues during embryonic development and in the adult brain. In transiently transfected cultures, mTIM promotes the nuclear localization of TIPIN. Immunoprecipitation experiments suggest that TIPIN is capable of regulating mTIM activity by disrupting the ability of mTIM to form homomultimeric complexes. Together, these results indicate that mTIM forms a functional complex with TIPIN, and provide a starting point from which to link mTim to biochemical pathways controlling vital cellular functions. (C) 2003 Elsevier Ltd. All rights reserved.