Transcription factors and aldolase B gene expression in microdissected renal proximal tubules and derived cell lines.
Renal expression of the aldolase B isoenzyme and transcription factors previously shown to regulate the aldolase B gene promoter in the liver were analyzed in whole kidney, microdissected tubules, and the two PKSV-PCT and PKSV-PR proximal tubule cell lines derived from transgenic mice. Aldolase B gene expression appeared restricted to the proximal tubule, the site where HNF1 alpha, HNF1 beta, C/EBP alpha, and DBP transcripts were also abundant. Compared to the liver, another organ synthesizing aldolase B, proximal tubules from the kidney were characterized by the absence of HNF3 and the presence of higher ratio of HNF1 beta/HNF1 alpha transcripts. The same features were conserved in both PKSV-PCT and PKSV-PR proximal tubule cell lines. Transactivation experiments in PKSV-PCT cultured cells showed that HNF1 alpha, C/EBP alpha, and DBP behave as transactivators of the 190-bp aldolase B gene promoter, and that HNF1 beta had a low transactivating efficiency. HNF1 beta, as well as HNF3, antagonized the HNF1 alpha-dependent transactivation of the aldolase B promoter. The fact that both HNF1 beta and HNF3 factors play similar negative roles by competitively binding close to or on the HNF1 site could suggest that, in proximal tubule renal cells, HNF1 beta has the same attenuator effect on the aldolase B gene promoter as HNF3 in hepatocytes. Thus, these results indicate that such models of established renal tubule cell lines, which have conserved the same features of parental cells, represent valuable tools for studies of the regulation of genes expressed in proximal tubules of the kidney.
Institut National de La Santé et de la Recherche Médicale (INSERM) U129, Institut Cochin de Génétique Moléculaire, Université René Descartes, Paris, France.
V
V
Vallet
M
M
Bens
B
B
Antoine
F
F
Levrat
L
L
Miquerol
A
A
Kahn
A
A
Vandewalle
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000502:physiology*; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D007668:Kidney / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation / Q000502:physiology
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D004247:DNA; D005634:Fructose-Bisphosphate Aldolase