Analysis of the oligomerization of myogenin and E2A products in vivo using a two-hybrid assay system

Tushar Chakraborty, James F. Martin, Eric N. Olson

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Members of the helix-loop-helix (HLH) family of proteins bind DNA and activate transcription as homo-and heterodimers. Myogenin is a muscle-specific HLH protein that binds DNA in vitro as a heterodimer with several widely expressed HLH proteins, such as the E2A gene products E12 and E47. We describe a method for detection of protein-protein interactions among HLH proteins in vivo in which dimerization through the HLH motif reconstructs a hybrid transcription factor containing the DNA-binding domain of yeast GAL4 linked to one HLH motif and the activation domain of VP-16 linked to another. We have used this assay to investigate whether myogenin forms homomeric or heteromeric complexes in vivo and to determine whether growth factors and oncogenes that inhibit myogenesis influence myogenin's ability to dimerize. The results show that myogenin heterodimerizes with E12 and E47 in vivo, but it does not homodimerize to a measurable extent. Peptide growth factors, as well as the immediate early gene products c-Jun, v-Fos, and c-Myc, inhibit the activity of myogenin through a mechanism independent of its association with E2A products.

Original languageEnglish (US)
Pages (from-to)17498-17501
Number of pages4
JournalJournal of Biological Chemistry
Volume267
Issue number25
StatePublished - Sep 5 1992

Fingerprint

Myogenin
Oligomerization
Two-Hybrid System Techniques
Assays
Helix-Loop-Helix Motifs
Proteins
DNA
Intercellular Signaling Peptides and Proteins
Genes
Immediate-Early Genes
Dimerization
Muscle Development
Etoposide
Transcription
Oncogenes
Yeast
Muscle
Transcription Factors
Yeasts
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Analysis of the oligomerization of myogenin and E2A products in vivo using a two-hybrid assay system. / Chakraborty, Tushar; Martin, James F.; Olson, Eric N.

In: Journal of Biological Chemistry, Vol. 267, No. 25, 05.09.1992, p. 17498-17501.

Research output: Contribution to journalArticle

@article{80f1e17a258347b39531ed1632c64662,
title = "Analysis of the oligomerization of myogenin and E2A products in vivo using a two-hybrid assay system",
abstract = "Members of the helix-loop-helix (HLH) family of proteins bind DNA and activate transcription as homo-and heterodimers. Myogenin is a muscle-specific HLH protein that binds DNA in vitro as a heterodimer with several widely expressed HLH proteins, such as the E2A gene products E12 and E47. We describe a method for detection of protein-protein interactions among HLH proteins in vivo in which dimerization through the HLH motif reconstructs a hybrid transcription factor containing the DNA-binding domain of yeast GAL4 linked to one HLH motif and the activation domain of VP-16 linked to another. We have used this assay to investigate whether myogenin forms homomeric or heteromeric complexes in vivo and to determine whether growth factors and oncogenes that inhibit myogenesis influence myogenin's ability to dimerize. The results show that myogenin heterodimerizes with E12 and E47 in vivo, but it does not homodimerize to a measurable extent. Peptide growth factors, as well as the immediate early gene products c-Jun, v-Fos, and c-Myc, inhibit the activity of myogenin through a mechanism independent of its association with E2A products.",
author = "Tushar Chakraborty and Martin, {James F.} and Olson, {Eric N.}",
year = "1992",
month = "9",
day = "5",
language = "English (US)",
volume = "267",
pages = "17498--17501",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "25",

}

TY - JOUR

T1 - Analysis of the oligomerization of myogenin and E2A products in vivo using a two-hybrid assay system

AU - Chakraborty, Tushar

AU - Martin, James F.

AU - Olson, Eric N.

PY - 1992/9/5

Y1 - 1992/9/5

N2 - Members of the helix-loop-helix (HLH) family of proteins bind DNA and activate transcription as homo-and heterodimers. Myogenin is a muscle-specific HLH protein that binds DNA in vitro as a heterodimer with several widely expressed HLH proteins, such as the E2A gene products E12 and E47. We describe a method for detection of protein-protein interactions among HLH proteins in vivo in which dimerization through the HLH motif reconstructs a hybrid transcription factor containing the DNA-binding domain of yeast GAL4 linked to one HLH motif and the activation domain of VP-16 linked to another. We have used this assay to investigate whether myogenin forms homomeric or heteromeric complexes in vivo and to determine whether growth factors and oncogenes that inhibit myogenesis influence myogenin's ability to dimerize. The results show that myogenin heterodimerizes with E12 and E47 in vivo, but it does not homodimerize to a measurable extent. Peptide growth factors, as well as the immediate early gene products c-Jun, v-Fos, and c-Myc, inhibit the activity of myogenin through a mechanism independent of its association with E2A products.

AB - Members of the helix-loop-helix (HLH) family of proteins bind DNA and activate transcription as homo-and heterodimers. Myogenin is a muscle-specific HLH protein that binds DNA in vitro as a heterodimer with several widely expressed HLH proteins, such as the E2A gene products E12 and E47. We describe a method for detection of protein-protein interactions among HLH proteins in vivo in which dimerization through the HLH motif reconstructs a hybrid transcription factor containing the DNA-binding domain of yeast GAL4 linked to one HLH motif and the activation domain of VP-16 linked to another. We have used this assay to investigate whether myogenin forms homomeric or heteromeric complexes in vivo and to determine whether growth factors and oncogenes that inhibit myogenesis influence myogenin's ability to dimerize. The results show that myogenin heterodimerizes with E12 and E47 in vivo, but it does not homodimerize to a measurable extent. Peptide growth factors, as well as the immediate early gene products c-Jun, v-Fos, and c-Myc, inhibit the activity of myogenin through a mechanism independent of its association with E2A products.

UR - http://www.scopus.com/inward/record.url?scp=0026713290&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026713290&partnerID=8YFLogxK

M3 - Article

C2 - 1325437

AN - SCOPUS:0026713290

VL - 267

SP - 17498

EP - 17501

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 25

ER -