title: RACK1 is involved in endothelial barrier regulation via its two novel interacting partners.
abstract: BACKGROUND
  RACK1, receptor for activated protein kinase C, serves as an anchor in multiple signaling pathways. TIMAP, TGF-β inhibited membrane-associated protein, is most abundant in endothelial cells with a regulatory effect on the endothelial barrier function. The interaction of TIMAP with protein phosphatase 1 (PP1cδ) was characterized, yet little is known about its further partners.
  
  
  RESULTS
  We identified two novel interacting partners of RACK1, namely, TGF-β inhibited membrane-associated protein, TIMAP, and farnesyl transferase. TIMAP is most abundant in endothelial cells where it is involved in the regulation of the barrier function. WD1-4 repeats of RACK1 were identified as critical regions of the interaction both with TIMAP and farnesyl transferase. Phosphorylation of TIMAP by activation of the cAMP/PKA pathway reduced the amount of TIMAP-RACK1 complex and enhanced translocation of TIMAP to the cell membrane in vascular endothelial cells. However, both membrane localization of TIMAP and transendothelial resistance were attenuated after RACK1 depletion. Farnesyl transferase, the enzyme responsible for prenylation and consequent membrane localization of TIMAP, is present in the RACK1-TIMAP complex in control cells, but it does not co-immunoprecipitate with TIMAP after RACK1 depletion.
  
  
  CONCLUSIONS
  Transient parallel linkage of TIMAP and farnesyl transferase to RACK1 could ensure prenylation and transport of TIMAP to the plasma membrane where it may attend in maintaining the endothelial barrier as a phosphatase regulator.
authors.affiliation: Department of Medical Chemistry, University of Debrecen Medical and Health Science Center, Egyetem tér 1, Debrecen, H 4032, Hungary. csortos@med.unideb.hu.
authors.forename: Anita
authors.initials: A
authors.lastname: Boratkó
authors.forename: Pál
authors.initials: P
authors.lastname: Gergely
authors.forename: Csilla
authors.initials: C
authors.lastname: Csortos