The cytoplasmic C-terminus of polycystin-1 increases cell proliferation in kidney epithelial cells through serum-activated and Ca(2+)-dependent pathway(s).
Polycystin-1 (PC1) is a large transmembrane protein important in renal differentiation and defective in most cases of autosomal dominant polycystic kidney disease (ADPKD), a common cause of renal failure in adults. Although the genetic basis of ADPKD has been elucidated, molecular and cellular mechanisms responsible for the dysregulation of epithelial cell growth in ADPKD cysts are still not well defined. We approached this issue by investigating the role of the carboxyl cytoplasmic domain of PC1 involved in signal transduction on the control of kidney cell proliferation. Therefore, we generated human HEK293 cells stably expressing the PC1 cytoplasmic tail as a membrane targeted TrkA-PC1 chimeric receptor protein (TrkPC1). We found that TrkPC1 increased cell proliferation through an increase in cytoplasmic Ca2+ levels and activation of PKC alpha, thereby upregulating D1 and D3 cyclin, downregulating p21waf1 and p27kip1 cyclin inhibitors, and thus inducing cell cycle progression from G0/G1 to the S phase. Interestingly, TrkPC1-dependent Ca2+ increase and PKC alpha activation are not constitutive, but require serum factor(s) as parallel component. In agreement with this observation, a significant increase in ERK1/2 phosphorylation was observed. Consistently, inhibitors specifically blocking either PKC alpha or ERK1/2 prevented the TrkPC1-dependent proliferation increase. NGF, the TrkA ligand, blocked this increase. We propose that in kidney epithelial cells the overexpression of PC1 C-terminus upregulates serum-evoked intracellular Ca2+ by counteracting the growth-suppression activity of endogenous PC1 and leading to an increase in cell proliferation.
Manzati
Elisa
E
Department of Biochemistry and Molecular Biology, University of Ferrara, Via Luigi Borsari 46, I-44100 Ferrara, Italy.
Aguiari
Gianluca
G
Banzi
Manuela
M
Manzati
Michele
M
Selvatici
Rita
R
Falzarano
Sofia
S
Maestri
Iva
I
Pinton
Paolo
P
Rizzuto
Rosario
R
del Senno
Laura
L
D001798:Blood Proteins / Q000378:metabolism* / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D018816:Genes, cdc / Q000502:physiology; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D007668:Kidney / Q000378:metabolism* / Q000503:physiopathology; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D016891:Polycystic Kidney, Autosomal Dominant / Q000378:metabolism / Q000503:physiopathology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D051571:Protein Kinase C-alpha; D017434:Protein Structure, Tertiary / Q000502:physiology; D011506:Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D020917:Receptor, trkA / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D050396:TRPP Cation Channels; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
D001798:Blood Proteins; D002118:Calcium; D018797:Cell Cycle Proteins; D048052:Mitogen-Activated Protein Kinase 3; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D011506:Proteins; D020917:Receptor, trkA; D011993:Recombinant Fusion Proteins; D050396:TRPP Cation Channels; C496628:PRKCA protein, human