Phosphorylation and activation of the DNA binding activity of purified Stat1 by the Janus protein-tyrosine kinases and the epidermal growth factor receptor

Frederick W Quelle; William Thierfelder; Bruce A Witthuhn; Bo Tang; Stanley Cohen; James N Ihle

doi:10.1074/jbc.270.35.20775

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Phosphorylation and activation of the DNA binding activity of purified Stat1 by the Janus protein-tyrosine kinases and the epidermal growth factor receptor

Journal article

Open access

Peer reviewed

Phosphorylation and activation of the DNA binding activity of purified Stat1 by the Janus protein-tyrosine kinases and the epidermal growth factor receptor

Frederick W Quelle, William Thierfelder, Bruce A Witthuhn, Bo Tang, Stanley Cohen and James N Ihle

The Journal of biological chemistry, Vol.270(35), pp.20775-20780

09/01/1995

DOI: 10.1074/jbc.270.35.20775

PMID: 7657660

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Abstract

The activation of Janus protein-tyrosine kinases (Jaks) and the subsequent phosphorylation and activation of latent signal transducers and activators of transcription (Stats) are common elements in signal transduction through the cytokine receptor superfamily. To assess the role and specificity of Jaks in Stat activation, we have utilized baculovirus expression systems to produce Stat1 and the Jaks. Co-expression of Stat1 with Tyk2, Jak1, or Jak2 resulted in the specific tyrosine phosphorylation of Stat1 at Tyr701, the residue phosphorylated in mammalian cells stimulated with interferon gamma. Alternatively, Stat1, purified to apparent homogeneity from insect cell extracts, was phosphorylated at Tyr701 in Jak immune complex kinase reactions. Phosphorylation of purified Stat1 was necessary and sufficient for the acquisition of DNA binding activity. The specificity in both systems was indicated by the inability of a Jak2 catalytically inactive mutant (Jak2-Glu882) or the Tec protein-tyrosine kinase to phosphorylate Stat1. However, immune complex-purified epidermal growth factor receptor was capable of phosphorylating purified Stat1 at Tyr701 and activating its DNA binding activity in in vitro reactions.

Phosphorylation

Transfection

Protein-Tyrosine Kinases - isolation & purification

Protein Biosynthesis

Protein-Tyrosine Kinases - metabolism

Humans

Molecular Sequence Data

Recombinant Proteins - biosynthesis

Trans-Activators - isolation & purification

Proteins - isolation & purification

Spodoptera

TYK2 Kinase

DNA-Binding Proteins - metabolism

Janus Kinase 2

Recombinant Proteins - isolation & purification

Peptide Mapping

Proto-Oncogene Proteins

Trans-Activators - biosynthesis

Protein-Tyrosine Kinases - biosynthesis

Recombinant Proteins - metabolism

Tyrosine

Amino Acid Sequence

Cell Line

Phosphopeptides - chemistry

ErbB Receptors - metabolism

DNA-Binding Proteins - isolation & purification

Mammals

Blotting, Western

Janus Kinase 1

Sequence Homology, Amino Acid

Animals

Proteins - metabolism

STAT1 Transcription Factor

Sheep

Trans-Activators - metabolism

Mice

DNA-Binding Proteins - biosynthesis

Phosphopeptides - isolation & purification

Details

Title: Subtitle: Phosphorylation and activation of the DNA binding activity of purified Stat1 by the Janus protein-tyrosine kinases and the epidermal growth factor receptor
Creators: Frederick W Quelle - Department of Biochemistry, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA
William Thierfelder
Bruce A Witthuhn
Bo Tang
Stanley Cohen
James N Ihle
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.270(35), pp.20775-20780
DOI: 10.1074/jbc.270.35.20775
PMID: 7657660
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: R01 DK/HL42932 / NIDDK NIH HHS P01 HL53749 / NHLBI NIH HHS P30 CA21765 / NCI NIH HHS
Language: English
Date published: 09/01/1995
Academic Unit: Neuroscience and Pharmacology; Internal Medicine
Record Identifier: 9984040219802771

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