Insulin regulation of a novel WD-40 repeat protein in adipocytes

B. D. Rodgers, M. A. Levine, M. Bernier, C. Montrose-Rafizadeh

    Research output: Contribution to journalArticle

    Abstract

    A 400 bp PCR product generated with degenerate primers derived from the glucagon-like peptide-1 receptor was used to screen a rat skeletal muscle cDNA library. The predicted amino acid sequence of the 978 bp open reading frame has a predicted Mr of 35 804, an estimated isoelectric point (pI) of 5.31 and contains seven WD-40 repeats, which are common to G-protein beta subunits (Gβ). Although chemically and structurally similar to Gβ subunits, the predicted amino acid sequence, when compared with the previously cloned Gβ isoforms, was found to be only 31-41% similar and thus was named Gβ-like (GβL, 'Gable'). Western blotting of whole-cell lysates and immunoprecipitates of membrane and cytosolic fractions of HEK 293 cells stably overexpressing a carboxy-terminal His-tagged GβL indicates that the protein is cytosolic and that it migrates at 42 kDa. A 4 kb transcript was detected in all tissues surveyed by northern blotting; however, an additional 2 kb transcript was detected in testis. Expression of GβL mRNA was highest in the brain and testis, followed by lung, heart, kidney, skeletal muscle, spleen and liver. In addition, reverse transcriptase/PCR showed that several other tissues and cell lines express GβL. The ubiquitous nature of the tissue expression pattern of GβL is similar to that of the insulin receptor, which suggests that insulin may influence GβL expression. Indeed, GβL protein and mRNA levels, in fully differentiated 3T3-L1 adipocytes, were upregulated by insulin in a concentration-dependent fashion. These changes were highly sensitive to insulin stimulation, being minimally affected by doses as low as 0.1 nM and maximally elevated by 1 nM doses. These data suggest that insulin regulates GβL production and imply that some of the actions of insulin may be mediated, in part, by this novel intracellular protein.

    Original languageEnglish (US)
    Pages (from-to)325-332
    Number of pages8
    JournalJournal of Endocrinology
    Volume168
    Issue number2
    DOIs
    StatePublished - 2001

    Fingerprint

    Adipocytes
    Insulin
    Proteins
    Testis
    Amino Acid Sequence
    Skeletal Muscle
    GTP-Binding Protein beta Subunits
    Messenger RNA
    HEK293 Cells
    Insulin Receptor
    Isoelectric Point
    Reverse Transcriptase Polymerase Chain Reaction
    Gene Library
    Northern Blotting
    Open Reading Frames
    Protein Isoforms
    Spleen
    Western Blotting
    WD40 Repeats
    Kidney

    ASJC Scopus subject areas

    • Endocrinology

    Cite this

    Rodgers, B. D., Levine, M. A., Bernier, M., & Montrose-Rafizadeh, C. (2001). Insulin regulation of a novel WD-40 repeat protein in adipocytes. Journal of Endocrinology, 168(2), 325-332. https://doi.org/10.1677/joe.0.1680325

    Insulin regulation of a novel WD-40 repeat protein in adipocytes. / Rodgers, B. D.; Levine, M. A.; Bernier, M.; Montrose-Rafizadeh, C.

    In: Journal of Endocrinology, Vol. 168, No. 2, 2001, p. 325-332.

    Research output: Contribution to journalArticle

    Rodgers, BD, Levine, MA, Bernier, M & Montrose-Rafizadeh, C 2001, 'Insulin regulation of a novel WD-40 repeat protein in adipocytes', Journal of Endocrinology, vol. 168, no. 2, pp. 325-332. https://doi.org/10.1677/joe.0.1680325
    Rodgers, B. D. ; Levine, M. A. ; Bernier, M. ; Montrose-Rafizadeh, C. / Insulin regulation of a novel WD-40 repeat protein in adipocytes. In: Journal of Endocrinology. 2001 ; Vol. 168, No. 2. pp. 325-332.
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