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Query: UNIPROT:P41181 (
collecting duct
)
5,183
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
The inner medullary
collecting duct
(IMCD) produces very high levels of endothelin-1 (ET-1) that acts as an autocrine inhibitor of IMCD Na
+
and water reabsorption. Recent studies suggest that IMCD ET-1 production is enhanced by extracellular hypertonicity as can occur during high salt intake. Although NFAT5 has been implicated in the IMCD ET-1 hypertonicity response, no studies in any cell type have identified NFAT5 as a transcriptional regulator of the
EDN1
gene; the current study examined this using a mouse IMCD cell line (IMCD3). Media hypertonicity increased IMCD3 ET-1 mRNA in a dose- and time-dependent manner associated with increased NFAT5 nuclear localization. Knockdown of NFAT5 using small-interfering RNA or by CRISPR/Cas9-mediated targeting of exon 4 of the NFAT5 gene reduced the ET-1 hypertonicity response. Chromatin immunoprecipitation using an NFAT5 antibody pulled down ET-1 promoter regions containing NFAT5 consensus binding sequences. Transfected ET-1 promoter reporter constructs revealed maximal hypertonicity-induced reporter activity in the proximal 1-kb region; mutation of the two NFAT5 consensus-binding sites in this region abolished hypertonicity-induced reporter activity. The 1-kb ET-1 promoter-reporter construct lost hypertonicity responsiveness when transfected in CRISPR/Cas9-induced NFAT5-deficient cells. In summary, these findings represent the first description that NFAT5 is a direct transcriptional regulator of the
EDN1
gene in IMCD cells and point to a potentially important mechanism by which body Na
+
homeostasis is maintained.
...
PMID:Identification of NFAT5 as a transcriptional regulator of the EDN1 gene in collecting duct. 3062 23
Endothelin-1 (ET-1) is a peptide hormone that functions as a vasoconstrictor in the vasculature, whereas in the
collecting duct
of the kidney it exerts blood pressure-lowering effects via natriuretic actions. Aberrant ET-1 signaling is associated with several pathological states including hypertension and chronic kidney disease. ET-1 expression is regulated largely through transcriptional control of the gene that encodes ET-1,
EDN1
. Here we report a long, non-coding RNA (lncRNA) that appears to be antisense to the
EDN1
gene, called
EDN1
-AS
. Because
EDN1
-AS
represents a potential novel mechanism to regulate ET-1 expression, we examined the regulation of
EDN1
-AS
expression and action. A putative glucocorticoid receptor response (GR) element upstream of the predicted
EDN1
-AS
transcription start site was identified using the ENCODE database and the UCSC genome browser. Two homozygous deletion clones of the element were generated using CRISPR/Cas9. This deletion resulted in a significant increase in the expression of
EDN1
-AS
, which was associated with increased secretion of ET-1 peptide from HK-2 cells (two-fold increase in KO cells vs. CNTL,
n
= 7,
P
< 0.05). Phenotypic characterization of these CRISPR clones revealed a difference in cell growth rates. Using a standard growth assay, we determined that the KO1 clone exhibited a three-fold increase in growth over 8 days compared to control cells (
n
= 4,
P
< 0.01) and the KO2 clone exhibited a two-fold increase (
n
= 4,
P
< 0.01). These results support a role for
EDN1
-AS
as a novel regulatory mechanism of ET-1 expression and cellular proliferation.
...
PMID:
EDN1-AS
, A Novel Long Non-coding RNA Regulating Endothelin-1 in Human Proximal Tubule Cells. 3223 91
