Protein O-Glucosyltransferases Rumi (RUMI)

  • Hideyuki Takeuchi
  • Robert S. Haltiwanger
Reference work entry


Protein O-glucosylation involves the addition of a β-linked glucose to the hydroxyl group of a serine residue within an epidermal growth factor-like (EGF) repeat. EGF repeats are small protein modules found in many cell surfaces and secreted proteins, defined by the presence of six conserved cysteine residues forming three disulfide bonds (Campbell and Bork 1993). O-Glucose was originally identified on EGF repeats from bovine clotting factors VII and IX in 1988 (Hase et al. 1988). Comparison of sequences surrounding the modified serine on these and other proteins led to the proposal of a consensus sequence for O-glucosylation, C1-X-S-X-P-C2, where C1 and C2 are the first and second conserved cysteines of the EGF repeat (Harris and Spellman 1993). Database searches using this consensus sequence reveal more than 40 proteins in mammals are predicted to be O-glucosylated (Fernandez-Valdivia et al. 2011;Rana et al. 2011), including all members of the Notch receptor family. Recent glycoproteomic site mapping data on mouse Notch1 reveal that all 16 of the predicted sites are modified, most at high stoichiometries, indicating that the consensus sequence accurately predicts modification (Rana et al. 2011). An additional site was found with alanine in place of proline (C1ASAAC2), suggesting that the consensus sequence can be expanded to C1-X-S-X-P/A-C2, adding a few additional predicted sites from database searches (Rana et al. 2011). The O-glucose is typically extended by two α3-linked xyloses to the trisaccharide, Xylα1-3Xylα1-3Glcβ1-O-Ser in mammals, although mono and disaccharide forms have also been seen (Moloney et al. 2000; Rana et al. 2011; Whitworth et al. 2010).


Notch Signaling Notch Activity Endoplasmic Reticulum Retention G189E Mutant CAP10 Domain 
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  1. Acar M, Jafar-Nejad H, Takeuchi H, Rajan A, Ibrani D, Rana NA, Pan H, Haltiwanger RS, Bellen HJ (2008) Rumi is a CAP10 domain glycosyltransferase that modifies Notch and is required for Notch signaling. Cell 132:247–258PubMedCentralPubMedCrossRefGoogle Scholar
  2. Busch C, Schomig K, Hofmann F, Aktories K (2000) Characterization of the catalytic domain of Clostridium novyi alpha-toxin. Infect Immun 68:6378–6383PubMedCentralPubMedCrossRefGoogle Scholar
  3. Campbell ID, Bork P (1993) Epidermal growth factor-like modules. Curr Opin Struct Biol 3:385–392CrossRefGoogle Scholar
  4. Chang YC, Kwon-Chung KJ (1999) Isolation, characterization, and localization of a capsule-associated gene, CAP10, of Cryptococcus neoformans. J Bacteriol 181:5636–5643PubMedCentralPubMedGoogle Scholar
  5. Fernandez-Valdivia R, Takeuchi H, Samarghandi A, Lopez M, Leonardi J, Haltiwanger RS, Jafar-Nejad H (2011) Regulation of mammalian Notch signaling and embryonic development by the protein O-glucosyltransferase Rumi. Development 138:1925–1934PubMedCrossRefGoogle Scholar
  6. Haines N, Irvine KD (2003) Glycosylation regulates Notch signaling. Nat Rev Mol Cell Biol 4:786–797PubMedCrossRefGoogle Scholar
  7. Harris RJ, Spellman MW (1993) O-linked fucose and other post-translational modifications unique to EGF modules. Glycobiology 3:219–224PubMedCrossRefGoogle Scholar
  8. Hase S, Kawabata S, Nishimura H, Takeya H, Sueyoshi T, Miyata T, Iwanaga S, Takao T, Shimonishi Y, Ikenaka T (1988) A new trisaccharide sugar chain linked to a serine residue in bovine blood coagulation factors VII and IX. J Biochem (Tokyo) 104:867–868Google Scholar
  9. Jafar-Nejad H, Leonardi J, Fernandez-Valdivia R (2010) Role of glycans and glycosyltransferases in the regulation of Notch signaling. Glycobiology 20:931–949PubMedCrossRefGoogle Scholar
  10. Kopan R, Ilagan MX (2009) The canonical Notch signaling pathway: unfolding the activation mechanism. Cell 137:216–233PubMedCentralPubMedCrossRefGoogle Scholar
  11. Lee TV, Takeuchi H, Jafar-Nejad H (2010) Regulation of notch signaling via O-glucosylation insights from Drosophila studies. Methods Enzymol 480:375–398PubMedCrossRefGoogle Scholar
  12. Leonardi J, Fernandez-Valdivia R, Li YD, Simcox AA, Jafar-Nejad H (2011) Multiple O-glucosylation sites on Notch function as a buffer against temperature-dependent loss of signaling. Development 138:3569–3578PubMedCrossRefGoogle Scholar
  13. Moloney DJ, Shair LH, Lu FM, Xia J, Locke R, Matta KL, Haltiwanger RS (2000) Mammalian Notch1 is modified with two unusual forms of O-linked glycosylation found on epidermal growth factor-like modules. J Biol Chem 275:9604–9611PubMedCrossRefGoogle Scholar
  14. Rampal R, Luther KB, Haltiwanger RS (2007) Notch signaling in normal and disease states: possible therapies related to glycosylation. Curr Mol Med 7:427–445PubMedCrossRefGoogle Scholar
  15. Rana NA, Haltiwanger RS (2011) Fringe benefits: functional and structural impacts of O-glycosylation on the extracellular domain of Notch receptors. Curr Opin Struct Biol 21:583–589PubMedCentralPubMedCrossRefGoogle Scholar
  16. Rana NA, Nita-Lazar A, Takeuchi H, Kakuda S, Luther KB, Haltiwanger RS (2011) O-glucose trisaccharide is present at high but variable stoichiometry at multiple sites on mouse Notch1. J Biol Chem 286:31623–31637PubMedCrossRefGoogle Scholar
  17. Sethi MK, Buettner FF, Krylov VB, Takeuchi H, Nifantiev NE, Haltiwanger RS, Gerardy-Schahn R, Bakker H (2010) Identification of glycosyltransferase 8 family members as xylosyltransferases acting on O-glucosylated notch epidermal growth factor repeats. J Biol Chem 285:1582–1586PubMedCrossRefGoogle Scholar
  18. Sethi MK, Buettner FF, Ashikov A, Krylov VB, Takeuchi H, Nifantiev NE, Haltiwanger RS, Gerardy-Schahn R, Bakker H (2012) Molecular cloning of a xylosyltransferase that transfers the second xylose to o-glucosylated epidermal growth factor repeats of notch. J Biol Chem 287:2739–2748PubMedCrossRefGoogle Scholar
  19. Shao L, Luo Y, Moloney DJ, Haltiwanger R (2002) O-Glycosylation of EGF repeats: identification and initial characterization of a UDP-glucose: protein O-glucosyltransferase. Glycobiology 12:763–770PubMedCrossRefGoogle Scholar
  20. Takeuchi H, Haltiwanger RS (2010) Role of glycosylation of Notch in development. Semin Cell Dev Biol 21:638–645PubMedCentralPubMedCrossRefGoogle Scholar
  21. Takeuchi H, Fernandez-Valdivia RC, Caswell DS, Nita-Lazar A, Rana NA, Garner TP, Weldeghiorghis TK, Macnaughtan MA, Jafar-Nejad H, Haltiwanger RS (2011) Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. Proc Natl Acad Sci U S A 108:16600–16605PubMedCentralPubMedCrossRefGoogle Scholar
  22. Takeuchi H, Kantharia J, Sethi MK, Bakker H, Haltiwanger RS (2012) Site-specific O-glucosylation of the epidermal growth factor-like (EGF) repeats of Notch: efficiency of glycosylation is affected by proper folding and amino acid sequence of individual EGF repeats. J Biol Chem 287:33934–33944PubMedCrossRefGoogle Scholar
  23. Teng Y, Liu Q, Ma J, Liu F, Han Z, Wang Y, Wang W (2006) Cloning, expression and characterization of a novel human CAP10-like gene hCLP46 from CD34(+) stem/progenitor cells. Gene 371:7–15PubMedCrossRefGoogle Scholar
  24. Wang Y, Chang N, Zhang T, Liu H, Ma W, Chu Q, Lai Q, Liu L, Wang W (2010) Overexpression of human CAP10- like protein 46 KD in T-acute lymphoblastic leukemia and acute myelogenous leukemia. Genet Test Mol Biomarkers 14:127–133PubMedCrossRefGoogle Scholar
  25. Whitworth GE, Zandberg WF, Clark T, Vocadlo DJ (2010) Mammalian Notch is modified by d-Xyl-alpha1-3-dXyl-alpha1-3-d-Glc-beta1-O-Ser: implementation of a method to study O-glucosylation. Glycobiology 20:287–299PubMedCrossRefGoogle Scholar

Copyright information

© Springer Japan 2014

Authors and Affiliations

  1. 1.Department of Biochemistry and Cell BiologyStony Brook UniversityStony BrookUSA

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