DOI: 10.1073/pnas.072072399. Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes

Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes

10.1073/pnas.072072399

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Increased flux of glucose through the hexosamine biosynthetic pathway (HSP) is believed to mediate hyperglycemia-induced insulin resistance in diabetes. The end product of the HSP, UDPβ- N -acetylglucosamine (GlcNAc), is a donor sugar nucleotide for complex glycosylation in the secretory pathway and for O-linked GlcNAc (O-GlcNAc) addition to nucleocytoplasmic proteins. Cycling of the O-GlcNAc posttranslational modification was blocked by pharmacological inhibition of O-GlcNAcase, the enzyme that catalyzes O-GlcNAc removal from proteins, with O -(2-acetamido-2-deoxy- d -glucopyranosylidene)amino- N -phenylcarbamate (PUGNAc). PUGNAc treatment increased levels of O-GlcNAc and caused insulin resistance in 3T3-L1 adipocytes. Insulin resistance induced through the HSP by glucosamine and chronic insulin treatment correlated with increased O-GlcNAc levels on nucleocytoplasmic proteins. Whereas insulin receptor autophosphorylation and insulin receptor substrate 2 tyrosine phosphorylation were not affected by PUGNAc inhibition of O-GlcNAcase, downstream phosphorylation of Akt at Thr-308 and glycogen synthase kinase 3β at Ser-9 was inhibited. PUGNAc-induced insulin resistance was associated with increased O-GlcNAc modification of several proteins including insulin receptor substrate 1 and β-catenin, two important effectors of insulin signaling. These results suggest that elevation of O-GlcNAc levels attenuate insulin signaling and contribute to the mechanism by which increased flux through the HSP leads to insulin resistance in adipocytes.

Bibliography

Vosseller, K., Wells, L., Lane, M. D., & Hart, G. W. (2002). Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes. Proceedings of the National Academy of Sciences, 99(8), 5313â5318.

Authors 4

Keith Vosseller (first)
Lance Wells (additional)
M. Daniel Lane (additional)
Gerald W. Hart (additional)

References 62 Referenced 401

H Yki-Jarvinen, V Koivisto Ann Clin Res 16, 74–83 (1984). / Ann Clin Res by Yki-Jarvinen H (1984)
10.2337/diacare.15.3.418
Y T Kruszynska, J M Olefsky J Invest Med 44, 413–428 (1996). / J Invest Med by Kruszynska Y T (1996)
10.1172/JCI112981
S R Hager, A L Jochen, R K Kalkhoff Am J Physiol 260, E353–E362 (1991). / Am J Physiol by Hager S R (1991)
10.1038/414813a
10.2337/diacare.13.6.610
10.1016/S0021-9258(19)67706-9
10.1172/JCI118876
10.2337/diab.45.8.1003
10.2337/diab.42.9.1333
10.1172/JCI118013
10.1007/BF00400719
10.1172/JCI118349
10.1172/JCI119390
10.1146/annurev.biochem.66.1.315
10.1096/fj.01-0094rev
10.1074/jbc.272.14.9308
10.1074/jbc.272.14.9316
10.1074/jbc.274.45.32015
10.1074/jbc.275.15.10983
10.1016/S0026-0495(98)90058-0
10.1007/978-1-4615-1885-3_10
10.1006/bbrc.1997.6110
10.1016/S0300-9084(01)01295-0
10.1126/science.1058714
10.1074/jbc.270.32.18961
10.1016/S0021-9258(18)82216-5
10.1074/jbc.271.46.28741
10.1021/bi0027480
10.1074/jbc.R000010200
10.1016/S0304-4165(99)00105-1
10.1210/endo-128-5-2415
10.1016/S0021-9258(17)32170-1
10.1074/jbc.M010420200
10.1074/jbc.M109656200
10.1074/jbc.273.6.3611
10.1074/jbc.272.12.7759
10.1073/pnas.87.1.251
10.1016/S0021-9258(19)85559-X
10.1006/abio.2001.5132
10.1016/S0021-9258(18)89409-1
Y Kido, J Nakae, D Accili J Clin Endocrinol Metab 86, 972–979 (2001). / J Clin Endocrinol Metab by Kido Y (2001)
10.1074/jbc.271.49.31372
10.1128/MCB.19.6.4008
10.1128/MCB.18.12.6971
10.1016/S0960-9822(06)00122-9
10.1042/bj2940625
10.1038/378785a0
10.1074/jbc.273.21.13150
10.1126/science.254.5029.274
M F White Recent Prog Horm Res 53, 119–138 (1998). / Recent Prog Horm Res by White M F (1998)
10.1046/j.1365-2443.1998.00198.x
10.1074/jbc.273.18.10823
10.2337/diabetes.49.6.981
10.1073/pnas.97.22.12222
10.2337/diabetes.48.8.1562
10.1074/jbc.M108594200
E Heart, W S Choi, C K Sung Am J Physiol 278, E103–E112 (2000). / Am J Physiol by Heart E (2000)
10.1038/sj.onc.1204064
10.1093/emboj/20.21.5999
10.1128/MCB.21.5.1633-1646.2001

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:45 a.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 5:10 p.m.)
Indexed	1 week ago (Aug. 20, 2025, 8:33 a.m.)
Issued	23 years, 4 months ago (April 16, 2002)
Published	23 years, 4 months ago (April 16, 2002)
Published Online	23 years, 4 months ago (April 16, 2002)
Published Print	23 years, 4 months ago (April 16, 2002)

Funders 0

None

BibTeX

@article{Vosseller_2002, title={Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes}, volume={99}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.072072399}, DOI={10.1073/pnas.072072399}, number={8}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Vosseller, Keith and Wells, Lance and Lane, M. Daniel and Hart, Gerald W.}, year={2002}, month=apr, pages={5313–5318} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        20
      ]
    ],
    "date-time": "2025-08-20T12:33:23Z",
    "timestamp": 1755693203180
  },
  "reference-count": 62,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "8",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2002,
        4,
        16
      ]
    ]
  },
  "abstract": "<jats:p>\n            Increased flux of glucose through the hexosamine biosynthetic pathway (HSP) is believed to mediate hyperglycemia-induced insulin resistance in diabetes. The end product of the HSP, UDP\u03b2-\n            <jats:italic>N</jats:italic>\n            -acetylglucosamine (GlcNAc), is a donor sugar nucleotide for complex glycosylation in the secretory pathway and for O-linked GlcNAc (O-GlcNAc) addition to nucleocytoplasmic proteins. Cycling of the O-GlcNAc posttranslational modification was blocked by pharmacological inhibition of O-GlcNAcase, the enzyme that catalyzes O-GlcNAc removal from proteins, with\n            <jats:italic>O</jats:italic>\n            -(2-acetamido-2-deoxy-\n            <jats:sc>d</jats:sc>\n            -glucopyranosylidene)amino-\n            <jats:italic>N</jats:italic>\n            -phenylcarbamate (PUGNAc). PUGNAc treatment increased levels of O-GlcNAc and caused insulin resistance in 3T3-L1 adipocytes. Insulin resistance induced through the HSP by glucosamine and chronic insulin treatment correlated with increased O-GlcNAc levels on nucleocytoplasmic proteins. Whereas insulin receptor autophosphorylation and insulin receptor substrate 2 tyrosine phosphorylation were not affected by PUGNAc inhibition of O-GlcNAcase, downstream phosphorylation of Akt at Thr-308 and glycogen synthase kinase 3\u03b2 at Ser-9 was inhibited. PUGNAc-induced insulin resistance was associated with increased O-GlcNAc modification of several proteins including insulin receptor substrate 1 and \u03b2-catenin, two important effectors of insulin signaling. These results suggest that elevation of O-GlcNAc levels attenuate insulin signaling and contribute to the mechanism by which increased flux through the HSP leads to insulin resistance in adipocytes.\n          </jats:p>",
  "DOI": "10.1073/pnas.072072399",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:45:09Z",
    "timestamp": 1027694709000
  },
  "page": "5313-5318",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 401,
  "title": "Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes",
  "prefix": "10.1073",
  "volume": "99",
  "author": [
    {
      "given": "Keith",
      "family": "Vosseller",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Department of Biological Chemistry, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205"
        }
      ]
    },
    {
      "given": "Lance",
      "family": "Wells",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biological Chemistry, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205"
        }
      ]
    },
    {
      "given": "M. Daniel",
      "family": "Lane",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biological Chemistry, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205"
        }
      ]
    },
    {
      "given": "Gerald W.",
      "family": "Hart",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Biological Chemistry, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        2002,
        4,
        16
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_3_1_2",
      "first-page": "74",
      "volume": "16",
      "author": "Yki-Jarvinen H",
      "year": "1984",
      "unstructured": "H Yki-Jarvinen, V Koivisto Ann Clin Res 16, 74\u201383 (1984).",
      "journal-title": "Ann Clin Res"
    },
    {
      "key": "e_1_3_3_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.2337/diacare.15.3.418"
    },
    {
      "key": "e_1_3_3_3_2",
      "first-page": "413",
      "volume": "44",
      "author": "Kruszynska Y T",
      "year": "1996",
      "unstructured": "Y T Kruszynska, J M Olefsky J Invest Med 44, 413\u2013428 (1996).",
      "journal-title": "J Invest Med"
    },
    {
      "key": "e_1_3_3_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI112981"
    },
    {
      "key": "e_1_3_3_5_2",
      "first-page": "E353",
      "volume": "260",
      "author": "Hager S R",
      "year": "1991",
      "unstructured": "S R Hager, A L Jochen, R K Kalkhoff Am J Physiol 260, E353\u2013E362 (1991).",
      "journal-title": "Am J Physiol"
    },
    {
      "key": "e_1_3_3_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/414813a"
    },
    {
      "key": "e_1_3_3_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.2337/diacare.13.6.610"
    },
    {
      "key": "e_1_3_3_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(19)67706-9"
    },
    {
      "key": "e_1_3_3_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI118876"
    },
    {
      "key": "e_1_3_3_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.2337/diab.45.8.1003"
    },
    {
      "key": "e_1_3_3_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.2337/diab.42.9.1333"
    },
    {
      "key": "e_1_3_3_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI118013"
    },
    {
      "key": "e_1_3_3_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00400719"
    },
    {
      "key": "e_1_3_3_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI118349"
    },
    {
      "key": "e_1_3_3_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI119390"
    },
    {
      "key": "e_1_3_3_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.biochem.66.1.315"
    },
    {
      "key": "e_1_3_3_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1096/fj.01-0094rev"
    },
    {
      "key": "e_1_3_3_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.14.9308"
    },
    {
      "key": "e_1_3_3_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.14.9316"
    },
    {
      "key": "e_1_3_3_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.274.45.32015"
    },
    {
      "key": "e_1_3_3_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.275.15.10983"
    },
    {
      "key": "e_1_3_3_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0026-0495(98)90058-0"
    },
    {
      "key": "e_1_3_3_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/978-1-4615-1885-3_10"
    },
    {
      "key": "e_1_3_3_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/bbrc.1997.6110"
    },
    {
      "key": "e_1_3_3_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0300-9084(01)01295-0"
    },
    {
      "key": "e_1_3_3_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1058714"
    },
    {
      "key": "e_1_3_3_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.32.18961"
    },
    {
      "key": "e_1_3_3_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(18)82216-5"
    },
    {
      "key": "e_1_3_3_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.46.28741"
    },
    {
      "key": "e_1_3_3_30_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi0027480"
    },
    {
      "key": "e_1_3_3_31_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.R000010200"
    },
    {
      "key": "e_1_3_3_32_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0304-4165(99)00105-1"
    },
    {
      "key": "e_1_3_3_33_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1210/endo-128-5-2415"
    },
    {
      "key": "e_1_3_3_34_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(17)32170-1"
    },
    {
      "key": "e_1_3_3_35_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M010420200"
    },
    {
      "key": "e_1_3_3_36_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M109656200"
    },
    {
      "key": "e_1_3_3_37_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.6.3611"
    },
    {
      "key": "e_1_3_3_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.12.7759"
    },
    {
      "key": "e_1_3_3_39_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.87.1.251"
    },
    {
      "key": "e_1_3_3_40_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(19)85559-X"
    },
    {
      "key": "e_1_3_3_41_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/abio.2001.5132"
    },
    {
      "key": "e_1_3_3_42_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(18)89409-1"
    },
    {
      "key": "e_1_3_3_43_2",
      "first-page": "972",
      "volume": "86",
      "author": "Kido Y",
      "year": "2001",
      "unstructured": "Y Kido, J Nakae, D Accili J Clin Endocrinol Metab 86, 972\u2013979 (2001).",
      "journal-title": "J Clin Endocrinol Metab"
    },
    {
      "key": "e_1_3_3_44_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.49.31372"
    },
    {
      "key": "e_1_3_3_45_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.19.6.4008"
    },
    {
      "key": "e_1_3_3_46_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.18.12.6971"
    },
    {
      "key": "e_1_3_3_47_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0960-9822(06)00122-9"
    },
    {
      "key": "e_1_3_3_48_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1042/bj2940625"
    },
    {
      "key": "e_1_3_3_49_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/378785a0"
    },
    {
      "key": "e_1_3_3_50_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.21.13150"
    },
    {
      "key": "e_1_3_3_51_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.254.5029.274"
    },
    {
      "key": "e_1_3_3_52_2",
      "first-page": "119",
      "volume": "53",
      "author": "White M F",
      "year": "1998",
      "unstructured": "M F White Recent Prog Horm Res 53, 119\u2013138 (1998).",
      "journal-title": "Recent Prog Horm Res"
    },
    {
      "key": "e_1_3_3_53_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2443.1998.00198.x"
    },
    {
      "key": "e_1_3_3_54_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.18.10823"
    },
    {
      "key": "e_1_3_3_55_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.2337/diabetes.49.6.981"
    },
    {
      "key": "e_1_3_3_56_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.97.22.12222"
    },
    {
      "key": "e_1_3_3_57_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.2337/diabetes.48.8.1562"
    },
    {
      "key": "e_1_3_3_58_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M108594200"
    },
    {
      "key": "e_1_3_3_59_2",
      "first-page": "E103",
      "volume": "278",
      "author": "Heart E",
      "year": "2000",
      "unstructured": "E Heart, W S Choi, C K Sung Am J Physiol 278, E103\u2013E112 (2000).",
      "journal-title": "Am J Physiol"
    },
    {
      "key": "e_1_3_3_60_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/sj.onc.1204064"
    },
    {
      "key": "e_1_3_3_61_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/20.21.5999"
    },
    {
      "key": "e_1_3_3_62_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.21.5.1633-1646.2001"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.072072399",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        12
      ]
    ],
    "date-time": "2022-04-12T21:10:09Z",
    "timestamp": 1649797809000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.072072399"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2002,
        4,
        16
      ]
    ]
  },
  "references-count": 62,
  "journal-issue": {
    "issue": "8",
    "published-print": {
      "date-parts": [
        [
          2002,
          4,
          16
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.072072399"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.072072399",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        2002,
        4,
        16
      ]
    ]
  },
  "assertion": [
    {
      "value": "2002-02-06",
      "order": 1,
      "name": "accepted",
      "label": "Accepted",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "2002-04-16",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}