Isolation of TAO1, a protein kinase that activates MEKs in stress- activated protein kinase cascades

Michele Hutchison, Kevin S. Berman, Melanie H. Cobb

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

Several components of the budding yeast pheromoneresponse pathway are conserved in mammalian mitogen-activated protein (MAP) kinase pathways. Thus, we used degenerate oligonucleotides derived from the sequence of the Saccharomyces cerevisiae protein kinase Ste20p to amplify related sequences from the rat. One of these sequences was used to clone a rat Ste20p homolog, which we called TAO1 for its one thousand and one amino acids. Northern analysis shows TAO1 is highly expressed in brain, as is a homolog TAO2. Recombinant TAO1 was expressed and purified from Sf9 cells. In vitro, it activated MAP/extracellular signal-regulated protein kinase (ERK) kinases (MEKs) 3, 4, and 6 of the stress-responsive MAP kinase pathways, but not MEK1 or 2 of the classical MAP kinase pathway. TAO1 activated MEK3 but not MEK4 or MEK6 in transfected cells. MEK3 coimmunoprecipitated with TAO1 when they were expressed in 293 cells. In addition, immunoreactive MEK3 endogenous to Sf9 cells copurified with TAO1 produced from a recombinant baculovirus. The activation of and binding to MEK3 by TAO1 implicates TAO1 in the regulation of the p38-containing stress-responsive MAP kinase pathway.

Original languageEnglish (US)
Pages (from-to)28625-28632
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number44
DOIs
StatePublished - Oct 30 1998

Fingerprint

Mitogen-Activated Protein Kinase Kinases
Heat-Shock Proteins
Mitogen-Activated Protein Kinases
Protein Kinases
Sf9 Cells
MAP Kinase Kinase Kinase 3
Saccharomyces cerevisiae Proteins
Rats
Saccharomycetales
Baculoviridae
Mitogen-Activated Protein Kinase 1
Extracellular Signal-Regulated MAP Kinases
Mitogens
Oligonucleotides
Clone Cells
Yeast
Amino Acids
Brain
Chemical activation
TAO1 protein kinase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Isolation of TAO1, a protein kinase that activates MEKs in stress- activated protein kinase cascades. / Hutchison, Michele; Berman, Kevin S.; Cobb, Melanie H.

In: Journal of Biological Chemistry, Vol. 273, No. 44, 30.10.1998, p. 28625-28632.

Research output: Contribution to journalArticle

@article{68cbe267dc1448f8b3a16adce641e10e,
title = "Isolation of TAO1, a protein kinase that activates MEKs in stress- activated protein kinase cascades",
abstract = "Several components of the budding yeast pheromoneresponse pathway are conserved in mammalian mitogen-activated protein (MAP) kinase pathways. Thus, we used degenerate oligonucleotides derived from the sequence of the Saccharomyces cerevisiae protein kinase Ste20p to amplify related sequences from the rat. One of these sequences was used to clone a rat Ste20p homolog, which we called TAO1 for its one thousand and one amino acids. Northern analysis shows TAO1 is highly expressed in brain, as is a homolog TAO2. Recombinant TAO1 was expressed and purified from Sf9 cells. In vitro, it activated MAP/extracellular signal-regulated protein kinase (ERK) kinases (MEKs) 3, 4, and 6 of the stress-responsive MAP kinase pathways, but not MEK1 or 2 of the classical MAP kinase pathway. TAO1 activated MEK3 but not MEK4 or MEK6 in transfected cells. MEK3 coimmunoprecipitated with TAO1 when they were expressed in 293 cells. In addition, immunoreactive MEK3 endogenous to Sf9 cells copurified with TAO1 produced from a recombinant baculovirus. The activation of and binding to MEK3 by TAO1 implicates TAO1 in the regulation of the p38-containing stress-responsive MAP kinase pathway.",
author = "Michele Hutchison and Berman, {Kevin S.} and Cobb, {Melanie H.}",
year = "1998",
month = "10",
day = "30",
doi = "10.1074/jbc.273.44.28625",
language = "English (US)",
volume = "273",
pages = "28625--28632",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "44",

}

TY - JOUR

T1 - Isolation of TAO1, a protein kinase that activates MEKs in stress- activated protein kinase cascades

AU - Hutchison, Michele

AU - Berman, Kevin S.

AU - Cobb, Melanie H.

PY - 1998/10/30

Y1 - 1998/10/30

N2 - Several components of the budding yeast pheromoneresponse pathway are conserved in mammalian mitogen-activated protein (MAP) kinase pathways. Thus, we used degenerate oligonucleotides derived from the sequence of the Saccharomyces cerevisiae protein kinase Ste20p to amplify related sequences from the rat. One of these sequences was used to clone a rat Ste20p homolog, which we called TAO1 for its one thousand and one amino acids. Northern analysis shows TAO1 is highly expressed in brain, as is a homolog TAO2. Recombinant TAO1 was expressed and purified from Sf9 cells. In vitro, it activated MAP/extracellular signal-regulated protein kinase (ERK) kinases (MEKs) 3, 4, and 6 of the stress-responsive MAP kinase pathways, but not MEK1 or 2 of the classical MAP kinase pathway. TAO1 activated MEK3 but not MEK4 or MEK6 in transfected cells. MEK3 coimmunoprecipitated with TAO1 when they were expressed in 293 cells. In addition, immunoreactive MEK3 endogenous to Sf9 cells copurified with TAO1 produced from a recombinant baculovirus. The activation of and binding to MEK3 by TAO1 implicates TAO1 in the regulation of the p38-containing stress-responsive MAP kinase pathway.

AB - Several components of the budding yeast pheromoneresponse pathway are conserved in mammalian mitogen-activated protein (MAP) kinase pathways. Thus, we used degenerate oligonucleotides derived from the sequence of the Saccharomyces cerevisiae protein kinase Ste20p to amplify related sequences from the rat. One of these sequences was used to clone a rat Ste20p homolog, which we called TAO1 for its one thousand and one amino acids. Northern analysis shows TAO1 is highly expressed in brain, as is a homolog TAO2. Recombinant TAO1 was expressed and purified from Sf9 cells. In vitro, it activated MAP/extracellular signal-regulated protein kinase (ERK) kinases (MEKs) 3, 4, and 6 of the stress-responsive MAP kinase pathways, but not MEK1 or 2 of the classical MAP kinase pathway. TAO1 activated MEK3 but not MEK4 or MEK6 in transfected cells. MEK3 coimmunoprecipitated with TAO1 when they were expressed in 293 cells. In addition, immunoreactive MEK3 endogenous to Sf9 cells copurified with TAO1 produced from a recombinant baculovirus. The activation of and binding to MEK3 by TAO1 implicates TAO1 in the regulation of the p38-containing stress-responsive MAP kinase pathway.

UR - http://www.scopus.com/inward/record.url?scp=0032582644&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032582644&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.44.28625

DO - 10.1074/jbc.273.44.28625

M3 - Article

C2 - 9786855

AN - SCOPUS:0032582644

VL - 273

SP - 28625

EP - 28632

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 44

ER -