Insulin stimulates and diabetes inhibits O-linked N-acetylglucosamine transferase and O-glycosylation of Sp1

Gipsy Majumdar, Jeremiah Wright, Paul Markowitz, Antonio Martinez-Hernandez, Rajendra Raghow, Solomon S. Solomon

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Abstract

Insulin stimulates both the biosynthesis of transcription factor Sp1 and its O-linked N-acetylglucosaminylation (O-GlcNAcylation), which promotes nuclear localization of Sp1 and its ability to transactivate calmodulin (CaM) gene transcription. To investigate this further, we incubated H-411E liver cells with insulin (10,000 μU/ml) and quantified the subcellular distribution of O-GlcNAc transferasse (OGT) and O-GlcNAc-modified Sp1. We also examined the phosphorylation of Sp1 using both Western blot and incorporation of 32P into Sp1. The results demonstrate that insulin, but not glucagon, stimulates OGT synthesis and enhances cytosolic staining of OGT (histochemical). Insulin increases O-GlcNAc-Sp1, which peaks at 30 min, followed by decline at 4 h. In contrast, insnlin initiates phosphorylation of Sp1 early, followed by a continued increase in phosphorylated Sp1 (PO4-Sp1) at 4 h. A reciprocal relationship between O-GlcNAc-Sp1 and PO4-Sp1 was observed. To explore the pathophysiological relevance, we localized OGT in liver sections from streptozototin (STZ)-induced diabetic rats. We observed that staining of OGT in STZ-induced diabetic rat liver is clearly diminished, but it was substantially restored after 6 days of insulin treatment. We conclude that insulin stimulates CaM gene transcription via a dynamic interplay between O-glycosylation and phosphorylation of Sp1 that modulates stability, mobility, subcellular compartmentalization, and activity.

Original languageEnglish (US)
Pages (from-to)3184-3192
Number of pages9
JournalDiabetes
Volume53
Issue number12
DOIs
StatePublished - Dec 1 2004

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Glycosylation
Insulin
Phosphorylation
Calmodulin
Liver
Sp1 Transcription Factor
Staining and Labeling
Glucagon
Genes
UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase
Western Blotting

All Science Journal Classification (ASJC) codes

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Insulin stimulates and diabetes inhibits O-linked N-acetylglucosamine transferase and O-glycosylation of Sp1. / Majumdar, Gipsy; Wright, Jeremiah; Markowitz, Paul; Martinez-Hernandez, Antonio; Raghow, Rajendra; Solomon, Solomon S.

In: Diabetes, Vol. 53, No. 12, 01.12.2004, p. 3184-3192.

Research output: Contribution to journalArticle

Majumdar, G, Wright, J, Markowitz, P, Martinez-Hernandez, A, Raghow, R & Solomon, SS 2004, 'Insulin stimulates and diabetes inhibits O-linked N-acetylglucosamine transferase and O-glycosylation of Sp1', Diabetes, vol. 53, no. 12, pp. 3184-3192. https://doi.org/10.2337/diabetes.53.12.3184
Majumdar, Gipsy ; Wright, Jeremiah ; Markowitz, Paul ; Martinez-Hernandez, Antonio ; Raghow, Rajendra ; Solomon, Solomon S. / Insulin stimulates and diabetes inhibits O-linked N-acetylglucosamine transferase and O-glycosylation of Sp1. In: Diabetes. 2004 ; Vol. 53, No. 12. pp. 3184-3192.
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