Hexosamines regulate leptin production in 3T3-L1 adipocytes through transcriptional mechanisms

Peili Zhang, Ellen S. Klenk, Marc A. Lazzaro, Lloyd B. Williams, Robert Considine

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Abstract

This study was undertaken to examine the regulation of leptin gene (LEP) transcription and leptin release by hexosamines in 3T3-L1 adipocytes. Glucosamine (1 mM), an intermediate in hexosamine biosynthesis, increased leptin release to 117.0 ± 7.3% (P = 0.0430; n = 9) and 134.6 ± 6.5% of the control value (P = 0.0367; n = 4) by 48 and 96 h, respectively. With 0.01 mM glucosamine, leptin release was increased to 120.0 ± 3.0% of the control value (P = 0.0069; n = 4) by 96 h of treatment. Glucose at 5 and 20 mM stimulated leptin release to 759 ± 227% and 1104 ± 316% of the control value over the 96-h culture period. Inhibition of hexosamine biosynthesis with 6-diazo-5-oxonorleucine (20 μM) reduced glucose-stimulated leptin release 13 ± 2.3% and 29.9 ± 6.6% at 24 and 96 h, respectively (n = 4; P < 0.05). A 24-h incubation in 5 mM glucose significantly increased (163.0 ± 19.3%; n = 7) the activity of a human LEP promoter electroporated into differentiated 3T3-L1 cells. Glucosamine (1 mM; 48 h) also increased LEP promoter activity 170.0 ± 13.0% (n = 5). Mutation of the three Sp1 binding sites in the LEP construct significantly reduced promoter activity. However, glucose (5 mM; 24 h) and glucosamine (1 mM; 48 h) increased the activity of the mutated promoter to 165 ± 40% (n = 8) and 143 ± 13% of the control value (n = 8). Glucosamine significantly increased O-glycosylation of Sp1 by 16.1 ± 4.5% (P = 0.0305; n = 3). These data demonstrate that glucose and hexosamines regulate leptin production through transcriptional mechanisms localized to the proximal portion of the LEP promoter. Hexosamine-mediated regulation of LEP gene expression does not depend on Sp1 binding to traditional sites on the promoter.

Original languageEnglish
Pages (from-to)99-106
Number of pages8
JournalEndocrinology
Volume143
Issue number1
DOIs
StatePublished - 2002

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Hexosamines
Leptin
Adipocytes
Glucosamine
Glucose
Genes
3T3-L1 Cells
Glycosylation
Human Activities

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Hexosamines regulate leptin production in 3T3-L1 adipocytes through transcriptional mechanisms. / Zhang, Peili; Klenk, Ellen S.; Lazzaro, Marc A.; Williams, Lloyd B.; Considine, Robert.

In: Endocrinology, Vol. 143, No. 1, 2002, p. 99-106.

Research output: Contribution to journalArticle

Zhang, Peili ; Klenk, Ellen S. ; Lazzaro, Marc A. ; Williams, Lloyd B. ; Considine, Robert. / Hexosamines regulate leptin production in 3T3-L1 adipocytes through transcriptional mechanisms. In: Endocrinology. 2002 ; Vol. 143, No. 1. pp. 99-106.
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abstract = "This study was undertaken to examine the regulation of leptin gene (LEP) transcription and leptin release by hexosamines in 3T3-L1 adipocytes. Glucosamine (1 mM), an intermediate in hexosamine biosynthesis, increased leptin release to 117.0 ± 7.3{\%} (P = 0.0430; n = 9) and 134.6 ± 6.5{\%} of the control value (P = 0.0367; n = 4) by 48 and 96 h, respectively. With 0.01 mM glucosamine, leptin release was increased to 120.0 ± 3.0{\%} of the control value (P = 0.0069; n = 4) by 96 h of treatment. Glucose at 5 and 20 mM stimulated leptin release to 759 ± 227{\%} and 1104 ± 316{\%} of the control value over the 96-h culture period. Inhibition of hexosamine biosynthesis with 6-diazo-5-oxonorleucine (20 μM) reduced glucose-stimulated leptin release 13 ± 2.3{\%} and 29.9 ± 6.6{\%} at 24 and 96 h, respectively (n = 4; P < 0.05). A 24-h incubation in 5 mM glucose significantly increased (163.0 ± 19.3{\%}; n = 7) the activity of a human LEP promoter electroporated into differentiated 3T3-L1 cells. Glucosamine (1 mM; 48 h) also increased LEP promoter activity 170.0 ± 13.0{\%} (n = 5). Mutation of the three Sp1 binding sites in the LEP construct significantly reduced promoter activity. However, glucose (5 mM; 24 h) and glucosamine (1 mM; 48 h) increased the activity of the mutated promoter to 165 ± 40{\%} (n = 8) and 143 ± 13{\%} of the control value (n = 8). Glucosamine significantly increased O-glycosylation of Sp1 by 16.1 ± 4.5{\%} (P = 0.0305; n = 3). These data demonstrate that glucose and hexosamines regulate leptin production through transcriptional mechanisms localized to the proximal portion of the LEP promoter. Hexosamine-mediated regulation of LEP gene expression does not depend on Sp1 binding to traditional sites on the promoter.",
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