Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase

Hideyuki Takeuchi, Rodrigo C. Fernańdez-Valdivia, Devin S. Caswell, Aleksandra Nita-Lazar, Nadia A. Rana, Thomas P. Garner, Thomas K. Weldeghiorghis, Megan A. Macnaughtan, Hamed Jafar-Nejad, Robert S. Haltiwanger

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52%), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDPglucose to an EGF repeat from human factor VII. Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( 50CASSPC 55) of factor VII EGF repeat. Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( 587CYSSPC 592), is also modified with either O-Xyl or O-glucose glycans in cells. Mutation of the second serine (S590A) causes a loss of O-Xyl but not O-glucose at this site. Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used.

Original languageEnglish (US)
Pages (from-to)16600-16605
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume108
Issue number40
DOIs
StatePublished - Oct 4 2011
Externally publishedYes

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Glucosyltransferases
Drosophila
Epidermal Growth Factor
Glucose
Serine
Factor VII
Proteins
Consensus Sequence
Uridine Diphosphate Glucose
Glycosyltransferases
Mutation
Uridine Diphosphate
Human Genome
Substrate Specificity
Endoplasmic Reticulum
Polysaccharides
UDP xylose-protein xylosyltransferase
Amino Acid Sequence
Clone Cells
Phenotype

Keywords

  • Development
  • Glycobiology
  • Mass spectrometry

ASJC Scopus subject areas

  • General

Cite this

Takeuchi, H., Fernańdez-Valdivia, R. C., Caswell, D. S., Nita-Lazar, A., Rana, N. A., Garner, T. P., ... Haltiwanger, R. S. (2011). Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. Proceedings of the National Academy of Sciences of the United States of America, 108(40), 16600-16605. https://doi.org/10.1073/pnas.1109696108

Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. / Takeuchi, Hideyuki; Fernańdez-Valdivia, Rodrigo C.; Caswell, Devin S.; Nita-Lazar, Aleksandra; Rana, Nadia A.; Garner, Thomas P.; Weldeghiorghis, Thomas K.; Macnaughtan, Megan A.; Jafar-Nejad, Hamed; Haltiwanger, Robert S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, No. 40, 04.10.2011, p. 16600-16605.

Research output: Contribution to journalArticle

Takeuchi, H, Fernańdez-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', Proceedings of the National Academy of Sciences of the United States of America, vol. 108, no. 40, pp. 16600-16605. https://doi.org/10.1073/pnas.1109696108
Takeuchi, Hideyuki ; Fernańdez-Valdivia, Rodrigo C. ; Caswell, Devin S. ; Nita-Lazar, Aleksandra ; Rana, Nadia A. ; Garner, Thomas P. ; Weldeghiorghis, Thomas K. ; Macnaughtan, Megan A. ; Jafar-Nejad, Hamed ; Haltiwanger, Robert S. / Rumi functions as both a protein O-glucosyltransferase and a protein O-xylosyltransferase. In: Proceedings of the National Academy of Sciences of the United States of America. 2011 ; Vol. 108, No. 40. pp. 16600-16605.
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abstract = "Mutations in rumi result in a temperature-sensitive loss of Notch signaling in Drosophila. Drosophila Rumi is a soluble, endoplasmic reticulum-retained protein with a CAP10 domain that functions as a protein O-glucosyltransferase. In human and mouse genomes, three potential Rumi homologues exist: one with a high degree of identity to Drosophila Rumi (52{\%}), and two others with lower degrees of identity but including a CAP10 domain (KDELC1 and KDELC2). Here we show that both mouse and human Rumi, but not KDELC1 or KDELC2, catalyze transfer of glucose from UDPglucose to an EGF repeat from human factor VII. Similarly, human Rumi, but not KDELC1 or KDELC2, rescues the Notch phenotypes in Drosophila rumi clones. During characterization of the Rumi enzymes, we noted that, in addition to protein O-glucosyltransferase activity, both mammalian and Drosophila Rumi also showed significant protein O-xylosyltransferase activity. Rumi transfers Xyl or glucose to serine 52 in the O-glucose consensus sequence ( 50CASSPC 55) of factor VII EGF repeat. Surprisingly, the second serine (S53) facilitates transfer of Xyl, but not glucose, to the EGF repeat by Rumi. EGF16 of mouse Notch2, which has a diserine motif in the consensus sequence ( 587CYSSPC 592), is also modified with either O-Xyl or O-glucose glycans in cells. Mutation of the second serine (S590A) causes a loss of O-Xyl but not O-glucose at this site. Altogether, our data establish dual substrate specificity for the glycosyltransferase Rumi and provide evidence that amino acid sequences of the recipient EGF repeat significantly influence which donor substrate (UDP-glucose or UDP-Xyl) is used.",
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AU - Nita-Lazar, Aleksandra

AU - Rana, Nadia A.

AU - Garner, Thomas P.

AU - Weldeghiorghis, Thomas K.

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