Human DRA Functions as a Sulfate Transporter in Sf9 Insect Cells

Byeon, Mee Kyeong; Frankel, Arthur E.; Papas, Takis S.; Henderson, Kelly W.; Schweinfest, Clifford W.

doi:10.1006/prep.1997.0809

Cited by 45 publications

(33 citation statements)

References 19 publications

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“…Consistent with our results, some previous studies on SLC26A5 and SLC26A6 have indicated that the N and C termini of these proteins are located in the cytosol (7,10,13,61). The location of the N terminus is important as previous reports have reported it to be proteolytically cleaved in vivo (65,66), raising the possibility that proteolytic cleavage may regulate transporter function. However, in this study the same banding pattern was observed in our Western blots regardless of whether an N-or C-terminal antibody was used (anti-c-Myc or anti-STAS domain; data not shown), suggesting that under our experimental conditions SLC26A3 is not undergoing any significant proteolysis.…”

Section: Discussionsupporting

confidence: 92%

Congenital Chloride-losing Diarrhea Causing Mutations in the STAS Domain Result in Misfolding and Mistrafficking of SLC26A3

Dorwart

Shcheynikov

Baker

et al. 2008

Journal of Biological Chemistry

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Congenital chloride-losing diarrhea (CLD) is a genetic disorder causing watery stool and dehydration. Mutations in SLC26A3 (solute carrier 26 family member 3), which functions as a coupled Cl ؊ /HCO 3 ؊ exchanger, cause CLD. SLC26A3 is a membrane protein predicted to contain 12 transmembranespanning ␣-helices and a C-terminal STAS (sulfate transporters and anti-sigma-factor) domain homologous to the bacterial anti-sigma-factor antagonists. The STAS domain is required for SLC26A3 Cl ؊ /HCO 3 ؊ exchange function and for the activation of cystic fibrosis transmembrane conductance regulator by SLC26A3. Here we investigate the molecular mechanism(s) by which four CLD-causing mutations (⌬Y526/7, I544N, I675/ 6ins, and G702Tins) in the STAS domain lead to disease. In a heterologous mammalian expression system biochemical, immunohistochemical, and ion transport experiments suggest that the four CLD mutations cause SLC26A3 transporter misfolding and/or mistrafficking. Expression studies with the isolated STAS domain suggest that the I675/6ins and G702Tins mutations disrupt the STAS domain directly, whereas limited proteolysis experiments suggest that the ⌬Y526/7 and I544N mutations affect a later step in the folding and/or trafficking pathway. The data suggest that these CLD-causing mutations cause disease by at least two distinct molecular mechanisms, both ultimately leading to loss of functional protein at the plasma membrane.

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Section: Discussionsupporting

confidence: 92%

Congenital Chloride-losing Diarrhea Causing Mutations in the STAS Domain Result in Misfolding and Mistrafficking of SLC26A3

Dorwart

Shcheynikov

Baker

et al. 2008

Journal of Biological Chemistry

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“…Another transporter from the same family is the protein known as "Down-Regulated in Adenoma" [DRA (SLC26A3)/Dra (Slc26a3)], which mediates Na + -independent SO 4 2-and oxalate uptake in mouse intestine (121). In transfected Sf9 insect cells (originate from insect ovarian tissue), human DRA mediated the DIDS-sensitive uptake of SO 4 2-and oxalate (122). In mice, Dra mRNA was only slightly expressed in the small intestine and strongly in the cecum and proximal colon, while in other organs the expression was below detection limits (121).…”

Section: Oxalate-handling Transporters and Their Role In Hyperoxalurimentioning

confidence: 99%

Oxalate: From the Environment to Kidney Stones

Brzica

Breljak

Burckhardt

et al. 2013

Archives of Industrial Hygiene and Toxicology

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Oxalate urolithiasis (nephrolithiasis) is the most frequent type of kidney stone disease. Epidemiological research has shown that urolithiasis is approximately twice as common in men as in women, but the underlying mechanism of this sex-related prevalence is unclear. Oxalate in the organism partially originate from food (exogenous oxalate) and largely as a metabolic end-product from numerous precursors generated mainly in the liver (endogenous oxalate). Oxalate concentrations in plasma and urine can be modifi ed by various foodstuffs, which can interact in positively or negatively by affecting oxalate absorption, excretion, and/or its metabolic pathways. Oxalate is mostly removed from blood by kidneys and partially via bile and intestinal excretion. In the kidneys, after reaching certain conditions, such as high tubular concentration and damaged integrity of the tubule epithelium, oxalate can precipitate and initiate the formation of stones. Recent studies have indicated the importance of the SoLute Carrier 26 (SLC26) family of membrane transporters for handling oxalate. Two members of this family [Sulfate Anion Transporter 1 (SAT-1; SLC26A1) and Chloride/Formate EXchanger (CFEX; SLC26A6)] may contribute to oxalate transport in the intestine, liver, and kidneys. Malfunction or absence of SAT-1 or CFEX has been associated with hyperoxaluria and urolithiasis. However, numerous questions regarding their roles in oxalate transport in the respective organs and male-prevalent urolithiasis, as well as the role of sex hormones in the expression of these transporters at the level of mRNA and protein, still remain to be answered.

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“…Two SO 4 2-transporters, named NaSi-1 (Slc13a1) and Sat-1 (Slc26a1), have been well studied and are suggested to be involved in SO 4 2-reabsorption. Slc13a1 is known to be a Na + /SO 4 2-cotransporter located on the apical or brush border membrane of the epithelial cells lining the renal proximal tubule and the small and large intestine of mammals (Besseghir and Roch-Ramel, 1987;Silberg et al, 1995;Byeon et al, 1998;Markovich, 2001). Slc26a1 is known to be a multifunctional anion exchanger located on the basolateral membrane of epithelial cells that have Slc13a1 on the apical side (Bissig et al, 1994;Karniski et al, 1998;Krick et al, 2009).…”

Section: +mentioning

confidence: 99%

Molecular physiology and functional morphology of SO42– excretion by the kidney of seawater-adapted eels

Watanabe

Takei

2011

Journal of Experimental Biology

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SUMMARY Marine teleosts actively excrete SO 42-and keep the plasma concentration of this ion much lower than that of environmental seawater (SW). We used the eel as a model to study the excretory mechanism of SO 4 2-because this euryhaline species changes SO 4 2-regulation drastically after transfer from freshwater (FW) to SW. Time-course studies showed that plasma SO 4 2-concentration decreased 3days after transfer of eels from FW to SW, while urine SO 4 2-concentration increased on 1day. Detailed analyses showed that urine SO 4 2-concentration increased linearly from 6h after SW transfer; however, this did not immediately translate to increased SO 4 2-excretion because the volume of urine was decreased. We identified five SO 4 2-transporters in the eel kidney. Three of these (Slc26a1, Slc26a6b and Slc26a6c) are expressed in both SW-and FW-acclimated eels while Slc26a6a and Slc13a1 are expressed in SW-acclimated eels and FW-acclimated eels, respectively. We showed that changes in Slc26a6a and Slc13a1 gene expression occurred 1-3days after SW transfer. In SW eel kidneys, immunohistochemistry using specific antisera against each transporter protein showed that Slc26a6a and Slc26a6c are localized on the apical membrane of the P1 segment of the proximal tubule, while Slc26a6b is localized on the apical membrane and Slc26a1 on the basolateral membrane of the P2 segment. The current study revealed complex molecular mechanisms of SO 4 2-excretion in the SW eel kidney that involve segment-specific localization of multiple Slc transporters in proximal tubules and modulation of their expression in different SO 4 2-environments. This precise regulatory mechanism may endow the eel with euryhalinity.

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Human DRA Functions as a Sulfate Transporter in Sf9 Insect Cells

Cited by 45 publications

References 19 publications

Congenital Chloride-losing Diarrhea Causing Mutations in the STAS Domain Result in Misfolding and Mistrafficking of SLC26A3

Congenital Chloride-losing Diarrhea Causing Mutations in the STAS Domain Result in Misfolding and Mistrafficking of SLC26A3

Oxalate: From the Environment to Kidney Stones

Molecular physiology and functional morphology of SO42– excretion by the kidney of seawater-adapted eels

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