Insulin activates a 70-kDa S6 kinase through serine/threonine-specific phosphorylation of the enzyme polypeptide

Price, D J; Gunsalus, J R; Avruch, J

doi:10.1073/pnas.87.20.7944

Title: Insulin activates a 70-kDa S6 kinase through serine/threonine-specific phosphorylation of the enzyme polypeptide

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Abstract

The dominant insulin-stimulated ribosomal protein S6 kinase activity was purified to near homogeneity from insulin-treated {sup 32}P-labeled rat H4 hepatoma cells and found to copurify with a 70-kDa {sup 32}P-labeled polypeptide. The dominant S6 kinase purified from livers of cycloheximide-treated rats is also a 70-kDa polypeptide. Antiserum raised against rat liver S6 kinase specifically immunoprecipitates the purified {sup 32}P-labeled H4 hepatoma insulin-stimulated S6 kinase. Immune complexes prepared from the cytosol of {sup 32}P-labeled H4 cells contain several {sup 32}P-labeled polypeptides. Insulin treatment increases the {sup 32}P content of the immunoprecipitated 70-kDa S6 kinase polypeptide 3- to 4-fold over basal levels. Tryptic peptide maps indicate that the insulin-stimulated S6 kinase purified from {sup 32}P-labeled H4 cells is phosphorylated at multiple sites distinct from those which participate in autophosphorylation in vitro. The S6 kinases purified from liver of cycloheximide-treated rat and H4 hepatoma insulin-stimulated enzyme are each completely deactivated by incubation with protein phosphatase type 2A in both autophosphorylating and 40S S6 phosphorylating activities. Thus insulin activates the 70-kDa S6 kinase by promoting phosphorylation of specific serine/threonine residues on the enzyme polypeptide, probably through activating an as-yet-unidentified serine/threonine protein kinase distinct from microtubule-associated protein 2 kinase.

Authors:

Price, D J; Gunsalus, J R; Avruch, J ^[1]

Harvard Medical School, Boston, MA (USA)

Publication Date:: Mon Oct 01 00:00:00 EDT 1990

OSTI Identifier:: 5802457

Resource Type:: Journal Article

Journal Name:: Proceedings of the National Academy of Sciences of the United States of America; (USA)

Additional Journal Information:: Journal Volume: 87:20; Journal ID: ISSN 0027-8424

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; INSULIN; BIOLOGICAL EFFECTS; PHOSPHOTRANSFERASES; ENZYME INDUCTION; POLYPEPTIDES; PHOSPHORYLATION; ATP; HEPATOMAS; PHOSPHORUS 32; RATS; SERINE; THREONINE; TUMOR CELLS; AMINO ACIDS; ANIMAL CELLS; ANIMALS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CARBOXYLIC ACIDS; CHEMICAL REACTIONS; DAYS LIVING RADIOISOTOPES; DISEASES; ENZYMES; GENE REGULATION; HORMONES; HYDROXY ACIDS; ISOTOPES; LIGHT NUCLEI; MAMMALS; NEOPLASMS; NUCLEI; NUCLEOTIDES; ODD-ODD NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; PEPTIDE HORMONES; PEPTIDES; PHOSPHORUS ISOTOPES; PHOSPHORUS-GROUP TRANSFERASES; PROTEINS; RADIOISOTOPES; RODENTS; TRANSFERASES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats


                    Price, D J, Gunsalus, J R, and Avruch, J. Insulin activates a 70-kDa S6 kinase through serine/threonine-specific phosphorylation of the enzyme polypeptide.  United States: N. p., 1990. 
        Web.  doi:10.1073/pnas.87.20.7944.

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                    Price, D J, Gunsalus, J R, & Avruch, J. Insulin activates a 70-kDa S6 kinase through serine/threonine-specific phosphorylation of the enzyme polypeptide.  United States.  https://doi.org/10.1073/pnas.87.20.7944

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                    Price, D J, Gunsalus, J R, and Avruch, J. 1990.  
        "Insulin activates a 70-kDa S6 kinase through serine/threonine-specific phosphorylation of the enzyme polypeptide".  United States.  https://doi.org/10.1073/pnas.87.20.7944.

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@article{osti_5802457,

  title        = {Insulin activates a 70-kDa S6 kinase through serine/threonine-specific phosphorylation of the enzyme polypeptide},

  author       = {Price, D J and Gunsalus, J R and Avruch, J},

  abstractNote = {The dominant insulin-stimulated ribosomal protein S6 kinase activity was purified to near homogeneity from insulin-treated {sup 32}P-labeled rat H4 hepatoma cells and found to copurify with a 70-kDa {sup 32}P-labeled polypeptide. The dominant S6 kinase purified from livers of cycloheximide-treated rats is also a 70-kDa polypeptide. Antiserum raised against rat liver S6 kinase specifically immunoprecipitates the purified {sup 32}P-labeled H4 hepatoma insulin-stimulated S6 kinase. Immune complexes prepared from the cytosol of {sup 32}P-labeled H4 cells contain several {sup 32}P-labeled polypeptides. Insulin treatment increases the {sup 32}P content of the immunoprecipitated 70-kDa S6 kinase polypeptide 3- to 4-fold over basal levels. Tryptic peptide maps indicate that the insulin-stimulated S6 kinase purified from {sup 32}P-labeled H4 cells is phosphorylated at multiple sites distinct from those which participate in autophosphorylation in vitro. The S6 kinases purified from liver of cycloheximide-treated rat and H4 hepatoma insulin-stimulated enzyme are each completely deactivated by incubation with protein phosphatase type 2A in both autophosphorylating and 40S S6 phosphorylating activities. Thus insulin activates the 70-kDa S6 kinase by promoting phosphorylation of specific serine/threonine residues on the enzyme polypeptide, probably through activating an as-yet-unidentified serine/threonine protein kinase distinct from microtubule-associated protein 2 kinase.},

  doi          = {10.1073/pnas.87.20.7944},

  url          = {https://www.osti.gov/biblio/5802457},
  journal      = {Proceedings of the National Academy of Sciences of the United States of America; (USA)},

  issn         = {0027-8424},

  number       = ,

  volume       = 87:20,

  place        = {United States},

  year         = {Mon Oct 01 00:00:00 EDT 1990},

  month        = {Mon Oct 01 00:00:00 EDT 1990}

}

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