The cytoplasmic tail of FPC antagonizes the full-length protein in the regulation of mTOR pathway

PLoS One. 2014 May 22;9(5):e95630. doi: 10.1371/journal.pone.0095630. eCollection 2014.

Abstract

FPC (fibrocystin or polyductin) is a single transmembrane receptor-like protein, responsible for the human autosomal recessive polycystic kidney disease (ARPKD). It was recently proposed that FPC undergoes a Notch-like cleavage and subsequently the cleaved carboxy(C)-terminal fragment translocates to the nucleus. To study the functions of the isolated C-tail, we expressed the intracellular domain of human FPC (hICD) in renal epithelial cells. By 3-dimensional (3D) tubulogenesis assay, we found that in contrast to tubule-like structures formed from control cells, hICD-expressing cells exclusively formed cyst-like structures. By western blotting, we showed that the Akt/mTOR pathway, indicated by increased phosphorylation of Akt at serine 473 and S6 kinase 1 at threonine 389, was constitutively activated in hICD-expressing cells, similar to that in FPC knockdown cells and ARPKD kidneys. Moreover, application of mTOR inhibitor rapamycin reduced the size of the cyst-like structures formed by hICD-expressing cells. Application of either LY294002 or wortmannin inhibited the activation of both S6K1 and Akt. Expression of full-length FPC inhibited the activation of S6 and S6 kinase whereas co-expression of hICD with full-length FPC antagonized the inhibitory effect of full-length FPC on mTOR. Taken together, we propose that FPC modulates the PI3K/Akt/mTOR pathway and the cleaved C-tail regulates the function of the full-length protein.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Child, Preschool
  • Enzyme Activation
  • Gene Knockdown Techniques
  • HEK293 Cells
  • Humans
  • Infant
  • Infant, Newborn
  • Kidney / cytology
  • Kidney / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphorylation
  • Polycystic Kidney, Autosomal Recessive / metabolism*
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins c-akt / metabolism
  • Receptors, Cell Surface / analysis
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Ribosomal Protein S6 Kinases / metabolism
  • Signal Transduction*
  • TOR Serine-Threonine Kinases / metabolism*

Substances

  • PKHD1 protein, human
  • Receptors, Cell Surface
  • Proto-Oncogene Proteins c-akt
  • Ribosomal Protein S6 Kinases
  • TOR Serine-Threonine Kinases