Calcitriol upregulates expression and activity of the 1b isoform of the plasma membrane calcium pump in immortalized distal kidney tubular cells.
The plasma membrane calcium pump (PMCA) is ubiquitously expressed in calcium transporting epithelia. PMCA is encoded by four distinct genes (PMCA1-4) which can be further posttranscriptionally modified. PMCA1b is the only isoform of PMCA1 expressed in kidney and intestine. Calcitriol upregulates PMCA protein expression and activity and PMCA1 mRNA expression in the intestine. Calcitriol has a similar effect on kidney distal tubule PMCA activity in vivo but the cellular basis for this effect has not been studied. PMCA expression in Madin-Darby bovine kidney (MDBK) cells, a distal kidney tubule cell line, was compared with a proximal tubule (LLC-PK1) and embryonic (HEK 293) kidney cell line. Only MDBK cells express PMCA1b mRNA and PMCA protein. In MDBK cells, calcitriol increased steady state expression of PMCA1b mRNA and protein and upregulated the functional activity of PMCA on calcium transport to a similar degree. Furthermore, calcitriol enhanced PMCA1b mRNA stability. These data are consistent with in vivo localization studies demonstrating the distal kidney tubule localization of PMCA protein. Furthermore, they indicate that calcitriol is an important regulator of PMCA activity in the kidney distal tubule by a pathway that includes translation and posttranscriptional modification of PMCA1b.
Glendenning
P
P
Department of Medicine, University of Western Australia, Nedlands, Western Australia, 6009, Australia. paulglen@cyllene.uwa.edu.au
Ratajczak
T
T
Dick
I M
IM
Prince
R L
RL
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D015139:Blotting, Southern; D015153:Blotting, Western; D064026:Calbindins; D002117:Calcitriol / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D027682:Cation Transport Proteins; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007684:Kidney Tubules / Q000166:cytology / Q000201:enzymology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D053499:Plasma Membrane Calcium-Transporting ATPases; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D020033:Protein Isoforms; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000096:biosynthesis; D019831:Sodium-Calcium Exchanger / Q000096:biosynthesis; D013997:Time Factors; D015854:Up-Regulation
C506952:ATP2B1 protein, human; D064026:Calbindins; D027682:Cation Transport Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D020033:Protein Isoforms; D012333:RNA, Messenger; D064030:S100 Calcium Binding Protein G; D019831:Sodium-Calcium Exchanger; C110374:sodium-calcium exchanger 1; D003609:Dactinomycin; D053499:Plasma Membrane Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases; D002117:Calcitriol