Thyroid cell responses to thyrotropin and 12-O-tetradecanoyl-phorbol-13-acetate: Translocation of protein kinase C and phosphorylation of thyroid cell polypeptide substrates

Akira Tanabe, Thor B. Nielsen, Sheela R Kadapakkam, James B. Field

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Not all of the effects of thyroid-stimulating hormone (TSH) on the thyroid are mediated by activation of the adenylate cyclase-cyclic AMP system, indicating that other control systems must also exist. Although a calcium-phospholipid-dependent protein kinase (protein kinase C) and specific substrates had been identified in thyroid tissue, their responsiveness to TSH and other stimulators has not been determined. In thyroid cells which had been preloaded with [32P]orthophosphate, TSH and 12-O-tetradecanoyl-phor-bol-13-acetate (TPA) increased the phosphorylation of a 33K polypeptide substrate within 5 min in a dose-dependent fashion. The effect was observed with 1 mU/ml TSH and 3 nm TPA and was maximal with 100 mU/ml TSH and 100 nm TPA. The biologically inactive analog of TPA, 4α-phorbol, had no effect. Isobutylmethylxanthine (IBMX) decreased the phosphorylation of the 33K polypeptide and inhibited the effect of TSH and TPA, indicating that the phosphorylation is not mediated by cyclic AMP. TSH and IBMX, but not TPA, augmented phosphorylation of a 38K polypeptide, suggesting involvement of cyclic AMP. In contrast TPA, but not TSH, increased the phosphorylation of 58K and 28K polypeptides. TSH, but not TPA or 4α-phorbol, elevated the cyclic AMP level of thyroid slices. Incubation of thyroid slices with TSH or TPA significantly decreased protein kinase C activity in the 100,000g cytosol fraction and increased it in an extract of plasma membranes. The effect was present within 5 min and was maximal by 30 min. The effect was observed with 100 mU/ml TSH or 1 nm TPA. The stimulation by TSH or TPA of protein kinase C and its translocation from the cytosol to the plasma membranes of thyroid tissue may provide another mechanism for control of thyroid Cell metabolism.

Original languageEnglish (US)
Pages (from-to)92-99
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume243
Issue number1
DOIs
StatePublished - Nov 15 1985
Externally publishedYes

Fingerprint

Phosphorylation
Tetradecanoylphorbol Acetate
Thyrotropin
Protein Kinase C
Thyroid Gland
Acetates
Peptides
Substrates
Cyclic AMP
Cell membranes
Cytosol
Acetate Kinase
Cell Membrane
Tissue
Adenylyl Cyclases
Metabolism
Chemical activation
Phosphates

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Thyroid cell responses to thyrotropin and 12-O-tetradecanoyl-phorbol-13-acetate : Translocation of protein kinase C and phosphorylation of thyroid cell polypeptide substrates. / Tanabe, Akira; Nielsen, Thor B.; Kadapakkam, Sheela R; Field, James B.

In: Archives of Biochemistry and Biophysics, Vol. 243, No. 1, 15.11.1985, p. 92-99.

Research output: Contribution to journalArticle

@article{87038104f27d4a20a9bad94e57097260,
title = "Thyroid cell responses to thyrotropin and 12-O-tetradecanoyl-phorbol-13-acetate: Translocation of protein kinase C and phosphorylation of thyroid cell polypeptide substrates",
abstract = "Not all of the effects of thyroid-stimulating hormone (TSH) on the thyroid are mediated by activation of the adenylate cyclase-cyclic AMP system, indicating that other control systems must also exist. Although a calcium-phospholipid-dependent protein kinase (protein kinase C) and specific substrates had been identified in thyroid tissue, their responsiveness to TSH and other stimulators has not been determined. In thyroid cells which had been preloaded with [32P]orthophosphate, TSH and 12-O-tetradecanoyl-phor-bol-13-acetate (TPA) increased the phosphorylation of a 33K polypeptide substrate within 5 min in a dose-dependent fashion. The effect was observed with 1 mU/ml TSH and 3 nm TPA and was maximal with 100 mU/ml TSH and 100 nm TPA. The biologically inactive analog of TPA, 4α-phorbol, had no effect. Isobutylmethylxanthine (IBMX) decreased the phosphorylation of the 33K polypeptide and inhibited the effect of TSH and TPA, indicating that the phosphorylation is not mediated by cyclic AMP. TSH and IBMX, but not TPA, augmented phosphorylation of a 38K polypeptide, suggesting involvement of cyclic AMP. In contrast TPA, but not TSH, increased the phosphorylation of 58K and 28K polypeptides. TSH, but not TPA or 4α-phorbol, elevated the cyclic AMP level of thyroid slices. Incubation of thyroid slices with TSH or TPA significantly decreased protein kinase C activity in the 100,000g cytosol fraction and increased it in an extract of plasma membranes. The effect was present within 5 min and was maximal by 30 min. The effect was observed with 100 mU/ml TSH or 1 nm TPA. The stimulation by TSH or TPA of protein kinase C and its translocation from the cytosol to the plasma membranes of thyroid tissue may provide another mechanism for control of thyroid Cell metabolism.",
author = "Akira Tanabe and Nielsen, {Thor B.} and Kadapakkam, {Sheela R} and Field, {James B.}",
year = "1985",
month = "11",
day = "15",
doi = "10.1016/0003-9861(85)90776-3",
language = "English (US)",
volume = "243",
pages = "92--99",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Thyroid cell responses to thyrotropin and 12-O-tetradecanoyl-phorbol-13-acetate

T2 - Translocation of protein kinase C and phosphorylation of thyroid cell polypeptide substrates

AU - Tanabe, Akira

AU - Nielsen, Thor B.

AU - Kadapakkam, Sheela R

AU - Field, James B.

PY - 1985/11/15

Y1 - 1985/11/15

N2 - Not all of the effects of thyroid-stimulating hormone (TSH) on the thyroid are mediated by activation of the adenylate cyclase-cyclic AMP system, indicating that other control systems must also exist. Although a calcium-phospholipid-dependent protein kinase (protein kinase C) and specific substrates had been identified in thyroid tissue, their responsiveness to TSH and other stimulators has not been determined. In thyroid cells which had been preloaded with [32P]orthophosphate, TSH and 12-O-tetradecanoyl-phor-bol-13-acetate (TPA) increased the phosphorylation of a 33K polypeptide substrate within 5 min in a dose-dependent fashion. The effect was observed with 1 mU/ml TSH and 3 nm TPA and was maximal with 100 mU/ml TSH and 100 nm TPA. The biologically inactive analog of TPA, 4α-phorbol, had no effect. Isobutylmethylxanthine (IBMX) decreased the phosphorylation of the 33K polypeptide and inhibited the effect of TSH and TPA, indicating that the phosphorylation is not mediated by cyclic AMP. TSH and IBMX, but not TPA, augmented phosphorylation of a 38K polypeptide, suggesting involvement of cyclic AMP. In contrast TPA, but not TSH, increased the phosphorylation of 58K and 28K polypeptides. TSH, but not TPA or 4α-phorbol, elevated the cyclic AMP level of thyroid slices. Incubation of thyroid slices with TSH or TPA significantly decreased protein kinase C activity in the 100,000g cytosol fraction and increased it in an extract of plasma membranes. The effect was present within 5 min and was maximal by 30 min. The effect was observed with 100 mU/ml TSH or 1 nm TPA. The stimulation by TSH or TPA of protein kinase C and its translocation from the cytosol to the plasma membranes of thyroid tissue may provide another mechanism for control of thyroid Cell metabolism.

AB - Not all of the effects of thyroid-stimulating hormone (TSH) on the thyroid are mediated by activation of the adenylate cyclase-cyclic AMP system, indicating that other control systems must also exist. Although a calcium-phospholipid-dependent protein kinase (protein kinase C) and specific substrates had been identified in thyroid tissue, their responsiveness to TSH and other stimulators has not been determined. In thyroid cells which had been preloaded with [32P]orthophosphate, TSH and 12-O-tetradecanoyl-phor-bol-13-acetate (TPA) increased the phosphorylation of a 33K polypeptide substrate within 5 min in a dose-dependent fashion. The effect was observed with 1 mU/ml TSH and 3 nm TPA and was maximal with 100 mU/ml TSH and 100 nm TPA. The biologically inactive analog of TPA, 4α-phorbol, had no effect. Isobutylmethylxanthine (IBMX) decreased the phosphorylation of the 33K polypeptide and inhibited the effect of TSH and TPA, indicating that the phosphorylation is not mediated by cyclic AMP. TSH and IBMX, but not TPA, augmented phosphorylation of a 38K polypeptide, suggesting involvement of cyclic AMP. In contrast TPA, but not TSH, increased the phosphorylation of 58K and 28K polypeptides. TSH, but not TPA or 4α-phorbol, elevated the cyclic AMP level of thyroid slices. Incubation of thyroid slices with TSH or TPA significantly decreased protein kinase C activity in the 100,000g cytosol fraction and increased it in an extract of plasma membranes. The effect was present within 5 min and was maximal by 30 min. The effect was observed with 100 mU/ml TSH or 1 nm TPA. The stimulation by TSH or TPA of protein kinase C and its translocation from the cytosol to the plasma membranes of thyroid tissue may provide another mechanism for control of thyroid Cell metabolism.

UR - http://www.scopus.com/inward/record.url?scp=0022403377&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022403377&partnerID=8YFLogxK

U2 - 10.1016/0003-9861(85)90776-3

DO - 10.1016/0003-9861(85)90776-3

M3 - Article

C2 - 2415065

AN - SCOPUS:0022403377

VL - 243

SP - 92

EP - 99

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 1

ER -