Interaction of MAP17 with NHERF3/4 induces translocation of the renal Na/Pi IIa transporter to the trans-Golgi.
The function of the NaPiIIa renal sodium-phosphate transporter is regulated through a complex network of interacting proteins. Several PDZ domain-containing proteins interact with its COOH terminus while the small membrane protein MAP17 interacts with its NH(2) end. To elucidate the function of MAP17, we identified its interacting proteins using both bacterial and mammalian two-hybrid systems. Several PDZ domain-containing proteins, including the four NHERF proteins, as well as NaPiIIa and NHE3, were found to bind to MAP17. The interactions of MAP17 with the NHERF proteins and with NaPiIIa were further analyzed in opossum kidney (OK) cells. Expression of MAP17 alone had no effect on the NaPiIIa apical membrane distribution, but coexpression of MAP17 and NHERF3 or NHERF4 induced internalization of NaPiIIa, MAP17, and the PDZ protein to the trans-Golgi network (TGN). This effect was not observed when MAP17 was cotransfected with NHERF1/2 proteins. Inhibition of protein kinase C (PKC) prevented expression of the three proteins in the TGN. Activation of PKC in OK cells transfected only with MAP17 induced complete degradation of MAP17 and NaPiIIa. When lysosomal degradation was prevented, both proteins accumulated in the TGN. When the dopamine D1-like receptor was activated with fenoldopam, both NaPiIIa and MAP17 also accumulated in the TGN. Finally, cotransfection of MAP17 and NHERF3 prevented the adaptive upregulation of phosphate transport activity in OK cells in response to low extracellular phosphate. Therefore, the interaction between MAP17, NHERF3/4, and NaPiIIa in the TGN could be an important intermediate or alternate path in the internalization of NaPiIIa.
Lanaspa
Miguel A
MA
Laboratory of Molecular Toxicology, Univ. of Zaragoza, Veterinary Faculty, Calle Miguel Servet 177, 50013 Zaragoza, Spain.
Giral
Héctor
H
Breusegem
Sophia Y
SY
Halaihel
Nabil
N
Baile
Goretti
G
Catalán
Julia
J
Carrodeguas
José A
JA
Barry
Nicholas P
NP
Levi
Moshe
M
Sorribas
Víctor
V
D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003513:Cycloheximide / Q000494:pharmacology; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004298:Dopamine / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D008566:Membranes / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D008871:Microvilli / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009693:Nucleic Acid Hybridization; D009893:Opossums / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000502:physiology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics / Q000502:physiology*; D050613:Sodium-Phosphate Cotransporter Proteins, Type IIa / Q000235:genetics / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection; D014178:Translocation, Genetic
D018076:DNA, Complementary; D008565:Membrane Proteins; C509767:Pdzk1ip1 protein, mouse; D010750:Phosphoproteins; D011500:Protein Synthesis Inhibitors; D017923:Sodium-Hydrogen Exchangers; D050613:Sodium-Phosphate Cotransporter Proteins, Type IIa; C093376:sodium-hydrogen exchanger regulatory factor; D003513:Cycloheximide; D005978:Glutathione; D004298:Dopamine