Thyroid hormone promotes serine phosphorylation of p53 by mitogen-activated protein kinase

A. Shih, H. Y. Lin, F. B. Davis, P. J. Davis

Research output: Contribution to journalArticle

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Abstract

L-Thyroxine (T 4) nongenomically promotes association of mitogen-activated protein kinase (MAPK) and thyroid hormone receptor TRβ1 (TR) in the cell nucleus, leading to serine phosphorylation of the receptor. The oncogene suppressor protein, p53, is serine phosphorylated by several kinases and is known to interact with TRβ1. We studied whether association of p53 and TR is modulated by T 4 and involves serine phosphorylation of p53 by MAPK. TR-replete 293T human kidney cells were incubated with a physiological concentration of T 4 for 10-90 min. Nuclear fractions were immunoprecipitated and the resulting proteins separated and immunoblotted for co-immunoprecipitated proteins. Activated MAPK immunoprecipitates of nuclei from T 4-treated cells accumulated p53 in a time-dependent manner; T 4 and T 4-agarose were more effective than T 3. T 4-induced nuclear complexing of p53 and MAPK was inhibited by PD 98059 (PD) and U0126, two MAPK kinase (MEK) inhibitors, and was absent in cells treated with MEK antisense oligonucleotide and in dominant negative Ras cells. T 4 also caused nuclear co-immunoprecipitation of TRβ1 and p53, an effect also inhibited by PD. Nuclear complexing of p53 and MAPK also occurred in HeLa cells, which lack functional TR. Constitutively activated MAPK caused phosphorylation of a recombinant p53-GST fusion protein in vitro; thus, p53 is a substrate for MAPK. An indicator of p53 transcriptional activity, accumulation of the immediate - early gene product, c-Jun, was inhibited by T 4. This T 4 effect was reversed by PD, indicating that the transcriptional activity of p53 was altered by T 4-directed MAPK-p53 interaction.

Original languageEnglish
Pages (from-to)2870-2878
Number of pages9
JournalBiochemistry
Volume40
Issue number9
DOIs
Publication statusPublished - Mar 6 2001
Externally publishedYes

Fingerprint

Phosphorylation
Mitogen-Activated Protein Kinases
Thyroid Hormones
Serine
Mitogen-Activated Protein Kinase Kinases
Proteins
Cells
Thyroid Hormone Receptors
Association reactions
Immediate-Early Genes
Antisense Oligonucleotides
Oncogene Proteins
Cell Nucleus
Thyroxine
HeLa Cells
Immunoprecipitation
Sepharose
Phosphotransferases
Fusion reactions
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Thyroid hormone promotes serine phosphorylation of p53 by mitogen-activated protein kinase. / Shih, A.; Lin, H. Y.; Davis, F. B.; Davis, P. J.

In: Biochemistry, Vol. 40, No. 9, 06.03.2001, p. 2870-2878.

Research output: Contribution to journalArticle

Shih, A. ; Lin, H. Y. ; Davis, F. B. ; Davis, P. J. / Thyroid hormone promotes serine phosphorylation of p53 by mitogen-activated protein kinase. In: Biochemistry. 2001 ; Vol. 40, No. 9. pp. 2870-2878.
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