Advertisement

BioMetals

, Volume 24, Issue 1, pp 159–170 | Cite as

Vesicular distribution of Secretory Pathway Ca2+-ATPase isoform 1 and a role in manganese detoxification in liver-derived polarized cells

  • Sharon Leitch
  • Mingye Feng
  • Sabina Muend
  • Lelita T. Braiterman
  • Ann L. Hubbard
  • Rajini RaoEmail author
Article

Abstract

Manganese is a trace element that is an essential co-factor in many enzymes critical to diverse biological pathways. However, excess Mn2+ leads to neurotoxicity, with psychiatric and motor dysfunction resembling parkinsonism. The liver is the main organ for Mn2+ detoxification by excretion into bile. Although many pathways of cellular Mn2+ uptake have been established, efflux mechanisms remain essentially undefined. In this study, we evaluated a potential role in Mn2+ detoxification by the Secretory Pathway Ca2+, Mn2+-ATPase in rat liver and a liver-derived cell model WIF-B that polarizes to distinct bile canalicular and sinusoidal domains in culture. Of two known isoforms, only secretory pathway Ca2+-ATPase isoform 1 (SPCA1) was expressed in liver and WIF-B cells. As previously observed in non-polarized cells, SPCA1 showed overlapping distribution with TGN38, consistent with Golgi/TGN localization. However, a prominent novel localization of SPCA1 to an endosomal population close to, but not on the basolateral membrane was also observed. This was confirmed by fractionation of rat liver homogenates which revealed dual distribution of SPCA1 to the Golgi/TGN and a fraction that included the early endosomal marker, EEA1. We suggest that this novel pool of endosomes may serve to sequester Mn2+ as it enters from the sinusoidal/basolateral domains. Isoform-specific partial knockdown of SPCA1 delayed cell growth and formation of canalicular domain by about 30% and diminished viability upon exposure to Mn2+. Conversely, overexpression of SPCA1 in HEK 293T cells conferred tolerance to Mn2+ toxicity. Taken together, our findings suggest a role for SPCA1 in Mn2+ detoxification in liver.

Keywords

Manganese Ca2+-ATPase Liver Fractionation Trans-Golgi network SPCA1 ATP2C1 

Notes

Acknowledgments

This work was supported by a grant from the National Institutes of Health (GM52414) to RR and a predoctoral award from the American Heart Association to MF. We thank Deepti Mohamalawari for excellent technical assistance in preliminary work.

Supplementary material

10534_2010_9384_MOESM1_ESM.jpg (176 kb)
Fig. S1 Selective role of SPCA1 in Mn2+ detoxification. (A) RT-PCR (left and middle panel) and Western blot (right panel) showing redundant expression and specific knockdown of SPCA1 and SPCA2 in HEK 293T cells. SPCA2 expression was evaluated by Western blot analysis of microsomes. (B) Cell viability, evaluated by MTT assay, of WIF-B cells after exposure to CuCl2. Note the lack of significant effect of SPCA1 knockdown. Data was normalized to growth in the absence of added Cu2+. (C) Viability of HEK 293T cells following exposure to Mn2+ or Cu2+. Note the lack of effect of SPCA1 knockdown. Data are representative of two independent experiments with similar results. (JPEG 177 kb)
10534_2010_9384_MOESM2_ESM.pdf (9 kb)
Supplementary material 2 (PDF 10 kb)
Supplemental Movie

Twenty-five 0.32 mm optical sections of WIF-B cells co-stained for SPCA1 (green) and HA321 (red) were obtained by confocal microscopy, as described under Experimental Procedures, and combined to generate a three-dimensional representation. Although labeling of SPCA1 appears close to the basolateral membrane, no significant co-localization with HA321 is observed (MOV 535 kb)

References

  1. Anderson JG, Cooney PT et al (2007) Inhibition of DAT function attenuates manganese accumulation in the globus pallidus. Environ Toxicol Pharmacol 23(2):179–184CrossRefPubMedGoogle Scholar
  2. Au C, Benedetto A et al (2008) Manganese transport in eukaryotes: the role of DMT1. Neurotoxicology 29(4):569–576CrossRefPubMedGoogle Scholar
  3. Au C, Benedetto A et al (2009) SMF-1, SMF-2 and SMF-3 DMT1 orthologues regulate and are regulated differentially by manganese levels in C. elegans. PLoS One 4(11):e7792CrossRefPubMedGoogle Scholar
  4. Ballatori N, Miles E et al (1987) Homeostatic control of manganese excretion in the neonatal rat. Am J Physiol 252(5 Pt 2):R842–R847PubMedGoogle Scholar
  5. Barceloux DG (1999) Manganese. J Toxicol Clin Toxicol 37(2):293–307CrossRefPubMedGoogle Scholar
  6. Beckman RA, Mildvan AS et al (1985) On the fidelity of DNA replication: manganese mutagenesis in vitro. Biochemistry 24(21):5810–5817CrossRefPubMedGoogle Scholar
  7. Bergeron JJ, Rachubinski RA et al (1982) Galactose transfer to endogenous acceptors within Golgi fractions of rat liver. J Cell Biol 92(1):139–146CrossRefPubMedGoogle Scholar
  8. Bolton EC, Mildvan AS et al (2002) Inhibition of reverse transcription in vivo by elevated manganese ion concentration. Mol Cell 9(4):879–889CrossRefPubMedGoogle Scholar
  9. Braiterman LT, Heffernan S et al (2008) JAM-A is both essential and inhibitory to development of hepatic polarity in WIF-B cells. Am J Physiol Gastrointest Liver Physiol 294(2):G576–G588CrossRefPubMedGoogle Scholar
  10. Butterworth RF (2010) Metal toxicity, liver disease and neurodegeneration. Neurotox Res 18(1):100–105CrossRefPubMedGoogle Scholar
  11. Devasahayam G, Burke DJ et al (2007) Golgi manganese transport is required for rapamycin signaling in Saccharomyces cerevisiae. Genetics 177(1):231–238CrossRefPubMedGoogle Scholar
  12. Duan X, Chang JH et al (2007) Disrupted-In-Schizophrenia 1 regulates integration of newly generated neurons in the adult brain. Cell 130(6):1146–1158CrossRefPubMedGoogle Scholar
  13. Durr G, Strayle J et al (1998) The medial-Golgi ion pump Pmr1 supplies the yeast secretory pathway with Ca2+ and Mn2+ required for glycosylation, sorting, and endoplasmic reticulum-associated protein degradation. Mol Biol Cell 9(5):1149–1162PubMedGoogle Scholar
  14. Faddy HM, Smart CE et al (2008) Localization of plasma membrane and secretory calcium pumps in the mammary gland. Biochem Biophys Res Commun 369(3):977–981CrossRefPubMedGoogle Scholar
  15. Feng M, Grice DM et al (2010) Store-independent activation of Orai1 by SPCA2 in mammary tumors. Cell 143(1):84–98CrossRefPubMedGoogle Scholar
  16. Finley JW (1998) Manganese uptake and release by cultured human hepato-carcinoma (Hep-G2) cells. Biol Trace Elem Res 64(1–3):101–118CrossRefPubMedGoogle Scholar
  17. Girijashanker K, He L et al (2008) Slc39a14 gene encodes ZIP14, a metal/bicarbonate symporter: similarities to the ZIP8 transporter. Mol Pharmacol 73(5):1413–1423CrossRefPubMedGoogle Scholar
  18. Jaag HM, Pogany J et al (2010) A host Ca2+/Mn2+ ion pump is a factor in the emergence of viral RNA recombinants. Cell Host Microbe 7(1):74–81CrossRefPubMedGoogle Scholar
  19. Kaiser J (2003) Manganese: a high-octane dispute. Science 300(5621):926–928CrossRefPubMedGoogle Scholar
  20. Kallay LM, McNickle A et al (2006) Scribble associates with two polarity proteins, Lgl2 and Vangl2, via distinct molecular domains. J Cell Biochem 99(2):647–664CrossRefPubMedGoogle Scholar
  21. Kanyo ZF, Scolnick LR et al (1996) Structure of a unique binuclear manganese cluster in arginase. Nature 383(6600):554–557CrossRefPubMedGoogle Scholar
  22. Klaassen CD (1974) Biliary excretion of manganese in rats, rabbits, and dogs. Toxicol Appl Pharmacol 29(3):458–468CrossRefPubMedGoogle Scholar
  23. Lapinskas PJ, Cunningham KW et al (1995) Mutations in PMR1 suppress oxidative damage in yeast cells lacking superoxide dismutase. Mol Cell Biol 15(3):1382–1388PubMedGoogle Scholar
  24. Lucchini RG, Martin CJ et al (2009) From manganism to manganese-induced parkinsonism: a conceptual model based on the evolution of exposure. Neuromolecular Med 11(4):311–321CrossRefPubMedGoogle Scholar
  25. Madejczyk MS, Boyer JL et al (2009) Hepatic uptake and biliary excretion of manganese in the little skate, Leucoraja erinacea. Comp Biochem Physiol C Toxicol Pharmacol 149(4):566–571CrossRefPubMedGoogle Scholar
  26. Maeda T, Sugiura R et al (2004) Pmr1, a P-type ATPase, and Pdt1, an Nramp homologue, cooperatively regulate cell morphogenesis in fission yeast: the importance of Mn2+ homeostasis. Genes Cells 9(1):71–82CrossRefPubMedGoogle Scholar
  27. Mandal D, Woolf TB et al (2000) Manganese selectivity of pmr1, the yeast secretory pathway ion pump, is defined by residue gln783 in transmembrane segment 6. Residue Asp778 is essential for cation transport. J Biol Chem 275(31):23933–23938CrossRefPubMedGoogle Scholar
  28. Mandal D, Rulli SJ et al (2003) Packing interactions between transmembrane helices alter ion selectivity of the yeast Golgi Ca2+/Mn2+-ATPase PMR1. J Biol Chem 278(37):35292–35298CrossRefPubMedGoogle Scholar
  29. McMillan G (2005) Is electric arc welding linked to manganism or Parkinson’s disease? Toxicol Rev 24(4):237–257CrossRefPubMedGoogle Scholar
  30. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63CrossRefPubMedGoogle Scholar
  31. Nyasae LK, Hubbard AL et al (2003) Transcytotic efflux from early endosomes is dependent on cholesterol and glycosphingolipids in polarized hepatic cells. Mol Biol Cell 14(7):2689–2705CrossRefPubMedGoogle Scholar
  32. Okunade GW, Miller ML et al (2007) Loss of the Atp2c1 secretory pathway Ca(2+)-ATPase (SPCA1) in mice causes Golgi stress, apoptosis, and midgestational death in homozygous embryos and squamous cell tumors in adult heterozygotes. J Biol Chem 282(36):26517–26527CrossRefPubMedGoogle Scholar
  33. Papavasiliou PS, Miller ST et al (1966) Role of liver in regulating distribution and excretion of manganese. Am J Physiol 211(1):211–216PubMedGoogle Scholar
  34. Shanks MR, Cassio D et al (1994) An improved polarized rat hepatoma hybrid cell line. Generation and comparison with its hepatoma relatives and hepatocytes in vivo. J Cell Sci 107(Pt 4):813–825PubMedGoogle Scholar
  35. Silva AC, Bock NA (2008) Manganese-enhanced MRI: an exceptional tool in translational neuroimaging. Schizophr Bull 34(4):595–604CrossRefPubMedGoogle Scholar
  36. Sorin A, Rosas G et al (1997) PMR1, a Ca2+-ATPase in yeast Golgi, has properties distinct from sarco/endoplasmic reticulum and plasma membrane calcium pumps. J Biol Chem 272(15):9895–9901CrossRefPubMedGoogle Scholar
  37. Ton VK, Mandal D et al (2002) Functional expression in yeast of the human secretory pathway Ca(2+), Mn(2+)-ATPase defective in Hailey–Hailey disease. J Biol Chem 277(8):6422–6427CrossRefPubMedGoogle Scholar
  38. Van Baelen K, Vanoevelen J et al (2001) The Golgi PMR1 P-type ATPase of Caenorhabditis elegans. Identification of the gene and demonstration of calcium and manganese transport. J Biol Chem 276(14):10683–10691CrossRefPubMedGoogle Scholar
  39. Vanoevelen J, Dode L et al (2005) The secretory pathway Ca2+/Mn2+-ATPase 2 is a Golgi-localized pump with high affinity for Ca2+ ions. J Biol Chem 280(24):22800–22808CrossRefPubMedGoogle Scholar
  40. Wedler FC, Ley BW (1994) Kinetic, ESR, and trapping evidence for in vivo binding of Mn(II) to glutamine synthetase in brain cells. Neurochem Res 19(2):139–144CrossRefPubMedGoogle Scholar
  41. Weisiger RA, Fridovich I (1973) Mitochondrial superoxide simutase. Site of synthesis and intramitochondrial localization. J Biol Chem 248(13):4793–4796PubMedGoogle Scholar
  42. Xiang M, Mohamalawari D et al (2005) A novel isoform of the secretory pathway Ca2+, Mn(2+)-ATPase, hSPCA2, has unusual properties and is expressed in the brain. J Biol Chem 280(12):11608–11614CrossRefPubMedGoogle Scholar
  43. Yin Z, Jiang H et al (2010) Ferroportin is a manganese-responsive protein that decreases manganese cytotoxicity and accumulation. J Neurochem 112(5):1190–1198CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Sharon Leitch
    • 1
  • Mingye Feng
    • 1
  • Sabina Muend
    • 1
  • Lelita T. Braiterman
    • 2
  • Ann L. Hubbard
    • 2
  • Rajini Rao
    • 1
    Email author
  1. 1.Department of PhysiologyJohns Hopkins University School of MedicineBaltimoreUSA
  2. 2.Cell BiologyJohns Hopkins University School of MedicineBaltimoreUSA

Personalised recommendations