Functional expression of a MAP kinase kinase in COS cells and recognition by an anti-STE7/byr1 antibody.

Abstract:

:Mitogen-activated protein (MAP) kinases p42mapk and p44mapk are activated by dual tyrosine and threonine phosphorylation in vivo. Both MAPKs are phosphorylated and activated in vitro by an activator recently identified as a protein-tyrosine/threonine kinase. We have isolated a putative cDNA for a MAP kinase kinase (MAPKK) and determined its structure [Proc. Natl. Acad. Sci. USA, in press]. The protein encoded by this cDNA shares sequence homology with two yeast protein kinases byr1 and STE7. We now report that stimulation with serum of COS cells expressing this shares sequence homology with two yeast protein kinases byr1 and STE7. We now report that stimulation with serum of COS cells expressing this protein amplifies MAPK activator activity markedly. The increased activity co-migrates during chromatography with the expressed 45 kDa protein, recognized by an anti-STE7/byr1 antibody, and is abrogated by treatment with phosphatase 2A. Thus, this cDNA encodes a functional MAPKK. The anti-STE7/byr1 antibody also recognized a 46 kDa COS cell protein that was resolved from the expressed MAPKK by anion-exchange chromatography. This immunoreactive protein co-eluted with endogenous MAPKK activity, suggesting identification of the immunoreactive band as monkey MAPKK.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Haystead CM,Wu J,Gregory P,Sturgill TW,Haystead TA

doi

10.1016/0014-5793(93)81481-e

subject

Has Abstract

pub_date

1993-02-08 00:00:00

pages

12-6

issue

1-2

eissn

0014-5793

issn

1873-3468

pii

001457939381481E

journal_volume

317

pub_type

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