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  • Title: Novel interaction between the human 5-HT7 receptor isoforms and PLAC-24/eIF3k.
    Author: De Martelaere K, Lintermans B, Haegeman G, Vanhoenacker P.
    Journal: Cell Signal; 2007 Feb; 19(2):278-88. PubMed ID: 16935469.
    Abstract:
    Three 5-HT(7) receptor isoforms are expressed in rat and man, which differ in the amino acid sequence of their C-terminus. Thus far, no changes have been observed in the pharmacological profile of all three isoforms. To further elucidate the signal transduction pathway specific for these receptor variants, we screened for possible interacting proteins of the C-terminus of the h5-HT(7(a)) variant in a human foetal brain cDNA library. Using a yeast two-hybrid assay, we isolated PLAC-24/eIF3k as a possible interacting candidate. The association of PLAC-24 with all three receptor variants was observed and further reconfirmed in vivo by co-immunoprecipitation of PLAC-24 with the full-length receptor isoforms in transfected COS-7 cells. Studies with different deletion mutants of the receptor showed that the interaction between PLAC-24 and the receptor is not restricted to the C-terminus of the receptor. PLAC-24/eIF3k consists of 3 domains: an N-terminal HAM domain, a central WH domain and a C-terminal tail. We generated different domain constructs of PLAC-24, which indicated that the HAM and WH domain both interact with the 5-HT(7(a)) receptor. Overexpression of PLAC-24 in HEK293 cells, stably expressing the h5-HT(7(a)) receptor, caused a threefold augmentation in the expression levels of the receptor. Co-localisation studies in COS-7 cells showed that PLAC-24 relocates from the nucleus and perinuclear sites towards the plasma membrane upon co-expression with the receptor. On the other hand, the expression of domain variants of PLAC-24 seems to block the translocation of the receptor towards the membrane. These observations suggest that PLAC-24 may play a role in the transport and the stabilisation of newly synthesised 5-HT(7) receptor towards the plasma membrane.
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