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Decreased sulfotransferase SULT1C2 gene expression in DPT-induced polycystic kidney
Kazunobu Sugimura1, Tomoaki Tanaka, Yoshihiko Tanaka
1Department of Urology, Osaka City University Medical School, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-8585, Japan. ksugimura@msic.med.osaka-cu.ac.jp
Background:
The pathogenesis of polycystic kidney disease (PKD) remains unclear despite the identification of the genes responsible for hereditary PKD. In this study, we investigated the alteration of gene expressions in an acquired PKD model induced by 2-amino-4,5-diphenylthiazole (DPT) using the differential display method.
Methods:
Kidney mRNA from a Sprague-Dawley rat fed with 1% DPT for 4 days and from a control rat was compared by the RT-PCR differential display method. Differentially expressed bands were re-amplified and subcloned. Using these subclones as probes, the changes in gene expressions were confirmed by Northern blot analysis. Subsequently, mouse kidney cDNA library was screened.
Results:
The isolated 1.5-kb cDNA contained an open reading frame encoding 296 amino acids, which shared 94.3% identity with rat SULT1C2 sulfotransferase, and was considered to be its mouse ortholog (GenBank Accession No. AY005469). Mouse SULT1C2 mRNA was abundant in the kidney and stomach among normal mouse tissues. The expression of SULT1C2 mRNA was decreased in the rat kidney after DPT feeding but not in the stomach. Mouse SULT1C2 was expressed successfully using pET plasmid vector and E. coli. The recombinant 34-kD protein was capable of catalyzing the sulfation of p-nitrophenol at a Km of 3.1 mmol/L, by utilizing 3'-phosphoadenosine 5'-phosphosulfate (PAPS) as the sulfate donor.
Conclusions:
Although the physiological substrate and function of SULT1C2 have yet to be elucidated, its down-regulation could be involved in the cystic changes of tubules by decreasing the sulfation of the tubular basement membrane components.
Insights
Down-regulation of mouse sulfotransferase SULT1C2 in acquired polycystic kidney disease (PKD) may contribute to tubular changes. This study identified SULT1C2's role in a rat model, revealing its decreased expression in affected kidneys.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Polycystic kidney disease (PKD) pathogenesis remains unclear, even with identified causative genes for hereditary forms.
- This study explores gene expression alterations in an acquired PKD model induced by 2-amino-4,5-diphenylthiazole (DPT).
Purpose of the Study:
- To investigate gene expression changes in a DPT-induced acquired polycystic kidney disease (PKD) model.
- To identify specific genes involved in the pathogenesis of acquired PKD.
Main Methods:
- Utilized RT-PCR differential display to compare kidney mRNA from DPT-fed rats and control rats.
- Confirmed differential gene expression using Northern blot analysis and screened a mouse kidney cDNA library.
- Successfully expressed and characterized the recombinant mouse SULT1C2 protein in E. coli.
Main Results:
- Identified a 1.5-kb cDNA as the mouse ortholog of rat SULT1C2 sulfotransferase (GenBank Accession No. AY005469).
- Found mouse SULT1C2 mRNA abundant in normal kidney and stomach tissues.
- Observed decreased SULT1C2 mRNA expression in the DPT-treated rat kidney but not in the stomach.
- Demonstrated the recombinant SULT1C2 protein's sulfotransferase activity on p-nitrophenol.
Conclusions:
- Down-regulation of SULT1C2 in the kidney may contribute to tubular cystic changes in acquired PKD.
- This decrease could result from reduced sulfation of tubular basement membrane components.
- Further research is needed to elucidate the physiological substrate and precise function of SULT1C2.