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Murine isoforms of UDP-GlcNAc 2-epimerase/ManNAc kinase: Secondary structures, expression profiles, and response to ManNAc therapy

Glycoconjugate Journal volume 30, pages 609–618 (2013)Cite this article

Abstract

The bifunctional enzyme UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE) catalyzes the first two committed steps in sialic acid synthesis. Non-allosteric GNE gene mutations cause the muscular disorder GNE myopathy (also known as hereditary inclusion body myopathy), whose exact pathology remains unknown. Increased knowledge of GNE regulation, including isoform regulation, may help elucidate the pathology of GNE myopathy. While eight mRNA transcripts encoding human GNE isoforms are described, we only identified two mouse Gne mRNA transcripts, encoding mGne1 and mGne2, homologous to human hGNE1 and hGNE2. Orthologs of the other human isoforms were not identified in mice. mGne1 appeared as the ubiquitously expressed, major mouse isoform. The mGne2 encoding transcript is differentially expressed and may act as a tissue-specific regulator of sialylation. mGne2 expression appeared significantly increased the first 2 days of life, possibly reflecting the high sialic acid demand during this period. Tissues of the knock-in Gne p.M712T mouse model had similar mGne transcript expression levels among genotypes, indicating no effect of the mutation on mRNA expression. However, upon treatment of these mice with N-acetylmannosamine (ManNAc, a Gne substrate, sialic acid precursor, and proposed therapy for GNE myopathy), Gne transcript expression, in particular mGne2, increased significantly, likely resulting in increased Gne enzymatic activities. This dual effect of ManNAc supplementation (increased flux through the sialic acid pathway and increased Gne activity) needs to be considered when treating GNE myopathy patients with ManNAc. In addition, the existence and expression of GNE isoforms needs consideration when designing other therapeutic strategies for GNE myopathy.

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Abbreviations

A. avenae :

Acidovorax avenae

B2M:

β2 microglobulin

BLAST:

Basic Local Alignment Search Tool

B. subtilis :

Bacillus subtilis

CDM:

Consensus Data Mining

CMP:

Cytidine monophosphate

DMRV:

Distal Myopathy with Rimmed Vacuoles

DYRK2:

Dual-specificity tyrosine phosphorylation regulated kinase 2

E:

Embryonic day

E. coli :

Escherichia coli

FDM:

Fragment Database Mining

GCNA:

N-acetyl-β-D-glucosaminidase

GlcNAc:

N-acetylglucosamine

GNE:

UDP-GlcNAc 2-epimerase/ManNAc kinase

GOR:

Garnier-Osguthorpe-Robson

H. arsenicoxydans :

Herminiimonas arsenicoxydans

HIBM:

Hereditary Inclusion Body Myopathy

His:

Histidine

H. sapiens :

Homo sapiens

Min:

Minutes

ManNAc:

N-acetylmannosamine

NCBI:

National Center for Biotechnology Information

Neu5Ac:

N-acetylneuraminic acid

OMIM:

Online Mendelian Inheritance in Man

P:

Postnatal day

P. carotovorum :

Pectobacterium carotovorum

PCR:

Polymerase chain reaction

Pdb:

Protein databank

PRPP:

Phosphoribosylpyrophosphate

PSIPRED:

Protein Structure Initiative Prediction

qPCR:

Quantitative real-time polymerase chain reaction

RNA:

Ribonucleic acid

SGK1:

Serum and glucocorticoid-induced kinase 1

S. gordonii :

Streptococcus gordonii

T. maritime :

Thermotoga maritima

UDP:

Uridine diphosphate

VAV1:

Vav-family member of guanine nucleotide exchange factors

References

  1. Hinderlich, S., Stasche, R., Zeitler, R., Reutter, W.: A bifunctional enzyme catalyzes the first two steps in N-acetylneuraminic acid biosynthesis of rat liver. Purification and characterization of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase. J. Biol. Chem. 272, 24313–24318 (1997)

    PubMed  Article  CAS  Google Scholar 

  2. Eisenberg, I., Avidan, N., Potikha, T., Hochner, H., Chen, M., Olender, T., Barash, M., Shemesh, M., Sadeh, M., Grabov-Nardini, G., Shmilevich, I., Friedmann, A., Karpati, G., Bradley, W.G., Baumbach, L., Lancet, D., Asher, E.B., Beckmann, J.S., Argov, Z., Mitrani-Rosenbaum, S.: The UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase gene is mutated in recessive hereditary inclusion body myopathy. Nat. Genet. 29, 83–87 (2001)

    PubMed  Article  CAS  Google Scholar 

  3. Kornfeld, S., Kornfeld, R., Neufeld, E.F., O'Brien, P.J.: The feedback control of sugar nucleotide biosynthesis in liver. Proc. Natl. Acad. Sci. U. S. A. 52, 371–379 (1964)

    PubMed  Article  CAS  Google Scholar 

  4. Seppala, R., Lehto, V.P., Gahl, W.A.: Mutations in the human UDP-N-acetylglucosamine 2-epimerase gene define the disease sialuria and the allosteric site of the enzyme. Am. J. Hum. Genet. 64, 1563–1569 (1999)

    PubMed  Article  CAS  Google Scholar 

  5. Chen, X., Varki, A.: Advances in the biology and chemistry of sialic acids. ACS Chem. Biol. 5, 163–176 (2010)

    PubMed  Article  CAS  Google Scholar 

  6. Varki, N.M., Varki, A.: Diversity in cell surface sialic acid presentations: implications for biology and disease. Lab. Invest. 87, 851–857 (2007)

    PubMed  Article  CAS  Google Scholar 

  7. Watts, G.D., Thorne, M., Kovach, M.J., Pestronk, A., Kimonis, V.E.: Clinical and genetic heterogeneity in chromosome 9p associated hereditary inclusion body myopathy: exclusion of GNE and three other candidate genes. Neuromuscul. Disord. 13, 559–567 (2003)

    PubMed  Article  Google Scholar 

  8. Reinke, S.O., Hinderlich, S.: Prediction of three different isoforms of the human UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase. FEBS Lett. 581, 3327–3331 (2007)

    PubMed  Article  CAS  Google Scholar 

  9. Yardeni, T., Choekyi, T., Jacobs, K., Ciccone, C., Patzel, K., Anikster, Y., Gahl, W.A., Kurochkina, N., Huizing, M.: Identification, Tissue Distribution, and Molecular Modeling of Novel Human Isoforms of the Key Enzyme in Sialic Acid Synthesis, UDP-GlcNAc 2-Epimerase/ManNAc Kinase. Biochemistry 50, 8914–8925 (2011)

    PubMed  Article  CAS  Google Scholar 

  10. Huizing, M., Krasnewich, D.M.: Hereditary inclusion body myopathy: a decade of progress. Biochim. Biophys. Acta 1792, 881–887 (2009)

    PubMed  Article  CAS  Google Scholar 

  11. Noguchi, S., Keira, Y., Murayama, K., Ogawa, M., Fujita, M., Kawahara, G., Oya, Y., Imazawa, M., Goto, Y., Hayashi, Y.K., Nonaka, I., Nishino, I.: Reduction of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase activity and sialylation in distal myopathy with rimmed vacuoles. J. Biol. Chem. 279, 11402–11407 (2004)

    PubMed  Article  CAS  Google Scholar 

  12. Schwarzkopf, M., Knobeloch, K.P., Rohde, E., Hinderlich, S., Wiechens, N., Lucka, L., Horak, I., Reutter, W., Horstkorte, R.: Sialylation is essential for early development in mice. Proc. Natl. Acad. Sci. U. S. A. 99, 5267–5270 (2002)

    PubMed  Article  CAS  Google Scholar 

  13. Malicdan, M.C., Noguchi, S., Nonaka, I., Hayashi, Y.K., Nishino, I.: A Gne knockout mouse expressing human GNE D176V mutation develops features similar to distal myopathy with rimmed vacuoles or hereditary inclusion body myopathy. Hum. Mol. Genet. 16, 2669–2682 (2007)

    PubMed  Article  CAS  Google Scholar 

  14. Galeano, B., Klootwijk, R., Manoli, I., Sun, M., Ciccone, C., Darvish, D., Starost, M.F., Zerfas, P.M., Hoffmann, V.J., Hoogstraten-Miller, S., Krasnewich, D.M., Gahl, W.A., Huizing, M.: Mutation in the key enzyme of sialic acid biosynthesis causes severe glomerular proteinuria and is rescued by N-acetylmannosamine. J. Clin. Invest. 117, 1585–1594 (2007)

    PubMed  Article  CAS  Google Scholar 

  15. Kakani, S., Yardeni, T., Poling, J., Ciccone, C., Niethamer, T., Klootwijk, E.D., Manoli, I., Darvish, D., Hoogstraten-Miller, S., Zerfas, P., Tian, E., Ten Hagen, K.G., Kopp, J.B., Gahl, W.A., Huizing, M.: The Gne M712T mouse as a model for human glomerulopathy. Am. J. Pathol. 180, 1431–1440 (2012)

    PubMed  Article  CAS  Google Scholar 

  16. Malicdan, M.C., Noguchi, S., Hayashi, Y.K., Nonaka, I., Nishino, I.: Prophylactic treatment with sialic acid metabolites precludes the development of the myopathic phenotype in the DMRV-hIBM mouse model. Nat. Med. 15, 690–695 (2009)

    PubMed  Article  CAS  Google Scholar 

  17. Reinke, S.O., Lehmer, G., Hinderlich, S., Reutter, W.: Regulation and pathophysiological implications of UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE) as the key enzyme of sialic acid biosynthesis. Biol. Chem. 390, 591–599 (2009)

    PubMed  Article  CAS  Google Scholar 

  18. Garnier, J., Osguthorpe, D.J., Robson, B.: Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J. Mol. Biol. 120, 97–120 (1978)

    PubMed  Article  CAS  Google Scholar 

  19. Kloczkowski, A., Ting, K.L., Jernigan, R.L., Garnier, J.: Combining the GOR V algorithm with evolutionary information for protein secondary structure prediction from amino acid sequence. Proteins 49, 154–166 (2002)

    PubMed  Article  CAS  Google Scholar 

  20. Cheng, H., Sen, T.Z., Kloczkowski, A., Margaritis, D., Jernigan, R.L.: Prediction of protein secondary structure by mining structural fragment database. Polymer. (Guildf). 46(4314–4321) (2005)

    Google Scholar 

  21. Sen, T.Z., Cheng, H., Kloczkowski, A., Jernigan, R.L.: A Consensus Data Mining secondary structure prediction by combining GOR V and Fragment Database Mining. Protein Sci. 15, 2499–2506 (2006)

    PubMed  Article  CAS  Google Scholar 

  22. Bryson, K., McGuffin, L.J., Marsden, R.L., Ward, J.J., Sodhi, J.S., Jones, D.T.: Protein structure prediction servers at University College London. Nucleic Acids Res. 33, W36–38 (2005)

    PubMed  Article  CAS  Google Scholar 

  23. Yardeni, T., Eckhaus, M., Morris, H.D., Huizing, M., Hoogstraten-Miller, S.: Retro-orbital injections in mice. Lab. Anim. (NY) 40(155–160) (2011)

    Google Scholar 

  24. Yardeni, T., Ciccone, C., Hoogstraten-Miller, S., Darvish, D., Anikster, Y., Maples, P.B., Jay, C.M., Gahl, W.A., Nemunaitis, J., Huizing, M.: A Non-Viral, GNE-Lipoplex Treatment to Correct Sialylation Defects in Gne-Mutant (M712T) Mice. In: Proceedings of the American Society for Gene & Cell Therapy, Annual Meeting, Washington, DC (2010)

  25. Schmittgen, T.D., Livak, K.J.: Analyzing real-time PCR data by the comparative C(T) method. Nat. Protoc. 3, 1101–1108 (2008)

    PubMed  Article  CAS  Google Scholar 

  26. Reinke, S.O., Eidenschink, C., Jay, C.M., Hinderlich, S.: Biochemical characterization of human and murine isoforms of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE). Glycoconj. J. 26, 415–422 (2009)

    PubMed  Article  CAS  Google Scholar 

  27. Eisenberg, I., Grabov-Nardini, G., Hochner, H., Korner, M., Sadeh, M., Bertorini, T., Bushby, K., Castellan, C., Felice, K., Mendell, J., Merlini, L., Shilling, C., Wirguin, I., Argov, Z., Mitrani-Rosenbaum, S.: Mutations spectrum of GNE in hereditary inclusion body myopathy sparing the quadriceps. Hum. Mutat. 21, 99 (2003)

    PubMed  Article  Google Scholar 

  28. Tong, Y., Tempel, W., Nedyalkova, L., Mackenzie, F., Park, H.W.: Crystal structure of the N-acetylmannosamine kinase domain of GNE. PLoS One 4, e7165 (2009)

    PubMed  Article  Google Scholar 

  29. Kurochkina, N., Yardeni, T., Huizing, M.: Molecular modeling of the bifunctional enzyme UDP-GlcNAc 2-epimerase/ManNAc kinase and predictions of structural effects of mutations associated with HIBM and sialuria. Glycobiology 20, 322–337 (2010)

    PubMed  Article  CAS  Google Scholar 

  30. Zhao, B., Lehr, R., Smallwood, A.M., Ho, T.F., Maley, K., Randall, T., Head, M.S., Koretke, K.K., Schnackenberg, C.G.: Crystal structure of the kinase domain of serum and glucocorticoid-regulated kinase 1 in complex with AMP PNP. Protein Sci. 16, 2761–2769 (2007)

    PubMed  Article  CAS  Google Scholar 

  31. Keppler, O.T., Hinderlich, S., Langner, J., Schwartz-Albiez, R., Reutter, W., Pawlita, M.: UDP-GlcNAc 2-epimerase: a regulator of cell surface sialylation. Science 284, 1372–1376 (1999)

    PubMed  Article  CAS  Google Scholar 

  32. Krause, S., Hinderlich, S., Amsili, S., Horstkorte, R., Wiendl, H., Argov, Z., Mitrani-Rosenbaum, S., Lochmuller, H.: Localization of UDP-GlcNAc 2-epimerase/ManAc kinase (GNE) in the Golgi complex and the nucleus of mammalian cells. Exp. Cell Res. 304, 365–379 (2005)

    PubMed  Article  CAS  Google Scholar 

  33. Ghaderi, D., Strauss, H.M., Reinke, S., Cirak, S., Reutter, W., Lucka, L., Hinderlich, S.: Evidence for dynamic interplay of different oligomeric states of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase by biophysical methods. J. Mol. Biol. 369, 746–758 (2007)

    PubMed  Article  CAS  Google Scholar 

  34. Yogev, O., Pines, O.: Dual targeting of mitochondrial proteins: mechanism, regulation and function. Biochim. Biophys. Acta 1808, 1012–1020 (2011)

    PubMed  Article  CAS  Google Scholar 

  35. Duncan, P.I., Raymond, F., Fuerholz, A., Sprenger, N.: Sialic acid utilisation and synthesis in the neonatal rat revisited. PLoS One 4, e8241 (2009)

    PubMed  Article  Google Scholar 

  36. Kontou, M., Weidemann, W., Bork, K., Horstkorte, R.: Beyond glycosylation: sialic acid precursors act as signaling molecules and are involved in cellular control of differentiation of PC12 cells. Biol. Chem. 390, 575–579 (2009)

    PubMed  Article  CAS  Google Scholar 

  37. Kean, E.L., Munster-Kuhnel, A.K., Gerardy-Schahn, R.: CMP-sialic acid synthetase of the nucleus. Biochim. Biophys. Acta 1673, 56–65 (2004)

    PubMed  Article  CAS  Google Scholar 

  38. Kershaw, D.B., Beck, S.G., Wharram, B.L., Wiggins, J.E., Goyal, M., Thomas, P.E., Wiggins, R.C.: Molecular cloning and characterization of human podocalyxin-like protein. Orthologous relationship to rabbit PCLP1 and rat podocalyxin. J. Biol. Chem. 272, 15708–15714 (1997)

    PubMed  Article  CAS  Google Scholar 

  39. Chou, H.H., Takematsu, H., Diaz, S., Iber, J., Nickerson, E., Wright, K.L., Muchmore, E.A., Nelson, D.L., Warren, S.T., Varki, A.: A mutation in human CMP-sialic acid hydroxylase occurred after the Homo-Pan divergence. Proc. Natl. Acad. Sci. U. S. A. 95, 11751–11756 (1998)

    PubMed  Article  CAS  Google Scholar 

  40. Nemunaitis, G., Maples, P.B., Jay, C., Gahl, W.A., Huizing, M., Poling, J., Tong, A.W., Phadke, A.P., Pappen, B.O., Bedell, C., Templeton, N.S., Kuhn, J., Senzer, N., Nemunaitis, J.: Hereditary inclusion body myopathy: single patient response to GNE gene Lipoplex therapy. J. Gene Med. 12, 403–412 (2010)

    PubMed  Article  CAS  Google Scholar 

  41. Nemunaitis, G., Jay, C.M., Maples, P.B., Gahl, W.A., Huizing, M., Yardeni, T., Tong, A.W., Phadke, A.P., Pappen, B.O., Bedell, C., Allen, H., Hernandez, C., Templeton, N.S., Kuhn, J., Senzer, N., Nemunaitis, J.: Hereditary Inclusion Body Myopathy: Single Patient Response to Intravenous Dosing of GNE Gene Lipoplex. Hum. Gene Ther. 22, 1331–1341 (2011)

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

We greatly appreciate the expert laboratory work of Dr. Shelley Hoogstraten-Miller, Katherine Berger, Katherine Patzel, and Adrian Astiz-Martinez. The authors thank Theresa Calhoun for her skilled assistance with mouse maintenance. We thank the HIBM Research group (Encino, CA) for providing the knock-in Gne p.M712T mouse model. This work was performed in partial fulfillment of the requirements for a PhD degree of T.Y., Sackler Faculty of Medicine, Tel Aviv University, Israel. This study was supported by the Intramural Research Program of the National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States (T.Y., K.J., T.K.N., C.C., W.A.G., and M.H.) and Research Funds of The School of Theoretical Modeling, Chevy Chase, Maryland, United States (N.K.).

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Affiliations

  1. Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, 10 Center Drive, MSC 1851 Bld 10, Rm 10C103, Bethesda, MD, 20892-1851, USA

    Tal Yardeni, Katherine Jacobs, Terren K. Niethamer, Carla Ciccone, William A. Gahl & Marjan Huizing

  2. Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel

    Tal Yardeni & Yair Anikster

  3. Department of Biophysics, The School of Theoretical Modeling, Chevy Chase, MD, 20825, USA

    Natalya Kurochkina

Authors
  1. Tal Yardeni
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  2. Katherine Jacobs
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  3. Terren K. Niethamer
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  4. Carla Ciccone
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  6. Natalya Kurochkina
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  7. William A. Gahl
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  8. Marjan Huizing
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Correspondence to Marjan Huizing.

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Yardeni, T., Jacobs, K., Niethamer, T.K. et al. Murine isoforms of UDP-GlcNAc 2-epimerase/ManNAc kinase: Secondary structures, expression profiles, and response to ManNAc therapy. Glycoconj J 30, 609–618 (2013). https://doi.org/10.1007/s10719-012-9459-1

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  • DOI: https://doi.org/10.1007/s10719-012-9459-1

Keywords

  • GNE myopathy
  • Isoforms
  • Mouse
  • UDP-GlcNAc 2-epimerase/ManNAc kinase