Abstract
Recent work using Drosophila melanogaster has shown that a protein O-glucosyltransferase called Rumi regulates Notch signaling. Studies on several alleles of rumi identified in a forward genetic screen indicated that Rumi is a temperature-sensitive regulator of Notch signaling in flies. Further genetic and rescue experiments demonstrated that Rumi is a general regulator of Drosophila Notch signaling. Biochemical analyses showed that Rumi adds glucose to specific EGF repeats in the extracellular domain of Notch receptor in the Drosophila S2 cell line. Furthermore, RNAi-mediated knockdown of Rumi in this cell line resulted in a severe decrease in the level of O-linked glucose on Notch. In this chapter, we discuss the genetic and biochemical methods used to determine the role of Rumi in the regulation of Notch signaling in flies.
Copyright (c) 2010 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Review
MeSH terms
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Animals
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Animals, Genetically Modified
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Drosophila / genetics*
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Drosophila / metabolism
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Drosophila Proteins / genetics
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Drosophila Proteins / isolation & purification
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Drosophila Proteins / metabolism*
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Drosophila Proteins / physiology*
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Enzyme Assays / methods
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Genetic Techniques*
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Glucosyltransferases / genetics
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Glucosyltransferases / isolation & purification
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Glucosyltransferases / metabolism*
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Glucosyltransferases / physiology
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Glycosylation
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Humans
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Protein Processing, Post-Translational / genetics
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Protein Processing, Post-Translational / physiology
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Receptors, Notch / genetics
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Receptors, Notch / isolation & purification
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Receptors, Notch / metabolism*
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Receptors, Notch / physiology*
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Signal Transduction / genetics
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Signal Transduction / physiology
Substances
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Drosophila Proteins
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N protein, Drosophila
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Receptors, Notch
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Glucosyltransferases
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Rumi protein, Drosophila