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PMID: 2108863 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Synergism in ternary complex formation between the dimeric glycoprotein p67SRF, polypeptide p62TCF and the c-fos serum response element.

The EMBO journal ·Vol. 9 ·No. 4 ·1990-04-00 ·Pages 1123-30

Schröter H, Mueller CG, Meese K, Nordheim A

Abstract

Transcriptional regulation of the c-fos proto-oncogene requires the serum response element (SRE) which is complexed by a multi-protein assembly observed both in vitro and in vivo. Two protein factors, p67SRF and p62TCF (previously called p62), are required to interact with the SRE for efficient induction of c-fos by serum. By quantitative band shift electrophoresis we measure at least a 50-fold increase in SRE affinity for p67SRF/p62TCF over p67SRF alone. Stoichiometrically we determine that the ternary complex with p62TCF involves p67SRF in dimeric form. We demonstrate that p67SRF is a glycosylated nuclear transcription factor carrying terminal N-acetylglucosamine (GlcNAc) as a post-translational modification. A proteolytic limit digestion product, approximately 13 kd in size, was generated from the p67SRF-SRE complex. This p67SRF-core domain binds SRE, can dimerize with p67SRF and is still able to form a ternary complex with p62TCF. Therefore, three functional activities can be ascribed to this small p67SRF-core domain: specific DNA binding, dimerization and interaction with p62TCF. We demonstrate that these functions map within the p67SRF core fragment containing the region between amino acids 93 and 222.

MeSH Terms
Base Sequence Chloroquine/pharmacology Gene Expression Regulation Glycosylation HeLa Cells/drug effects,metabolism Humans Kinetics Macromolecular Substances Molecular Sequence Data Nuclear Proteins/physiology Oligonucleotide Probes Plasmids Polymerase Chain Reaction Protein-Tyrosine Kinases/genetics Proto-Oncogene Mas Proto-Oncogene Proteins/genetics Proto-Oncogene Proteins c-fos Proto-Oncogenes Serum Response Factor Transcription Factors/metabolism Transcription, Genetic
Chemicals
MAS1 protein, human Macromolecular Substances Nuclear Proteins Oligonucleotide Probes Proto-Oncogene Mas Proto-Oncogene Proteins Proto-Oncogene Proteins c-fos Serum Response Factor Transcription Factors Chloroquine Protein-Tyrosine Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Schröter H
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, FRG.
Mueller C G
Meese K
Nordheim A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-04-00
Pages
1123-30
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551787
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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