HASH-1 and E2-2 are expressed in human neuroblastoma cells and form a functional complex

Biochem Biophys Res Commun. 2000 Jul 21;274(1):22-31. doi: 10.1006/bbrc.2000.3090.

Abstract

The basic helix-loop-helix (bHLH) transcription factor mammalian achaete-scute homolog-1 (MASH-1 in mouse and HASH-1 in human) is essential for proper development of olfactory and most peripheral autonomic neurons, and for the formation of distinct neuronal circuits within the central nervous system. We have previously shown that HASH-1 is expressed in neuroblastoma tumors and cell lines, and in this study we have used the yeast two-hybrid system to isolate HASH-1 interacting proteins from a human neuroblastoma cDNA library. Two of the isolated clones contained cDNA from the E2-2 gene (also known as ITF2/SEF2-1). We show that E2-2 interacts with HASH-1 in both yeast and mammalian cells. The HASH-1/E2-2 complex binds an E-box (CACCTG) in vitro, and transactivates an E-box containing reporter construct in vivo. Furthermore, E2-2 seems to be one of the major HASH-1 interacting proteins in extracts from neuroblastoma cells. In conclusion, E2-2 forms a functional complex with HASH-1, and might therefore be involved in the development of specific parts of the central and peripheral nervous systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
  • Basic Helix-Loop-Helix Transcription Factors
  • Blotting, Northern
  • Blotting, Western
  • Cell Differentiation
  • DNA, Complementary / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • Gene Library
  • Glutathione Transferase / metabolism
  • HeLa Cells
  • Humans
  • Luciferases / metabolism
  • Nerve Tissue Proteins*
  • Neuroblastoma / genetics
  • Neuroblastoma / metabolism*
  • Plasmids / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / metabolism
  • TCF Transcription Factors
  • Time Factors
  • Trans-Activators / chemistry
  • Trans-Activators / metabolism*
  • Transcription Factor 4
  • Transcription Factor 7-Like 2 Protein
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism*
  • Transcriptional Activation
  • Transfection
  • Tumor Cells, Cultured
  • Two-Hybrid System Techniques
  • Yeasts / metabolism

Substances

  • ASCL1 protein, human
  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
  • Basic Helix-Loop-Helix Transcription Factors
  • DNA, Complementary
  • DNA-Binding Proteins
  • Nerve Tissue Proteins
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • TCF Transcription Factors
  • TCF4 protein, human
  • TCF7L2 protein, human
  • Tcf4 protein, mouse
  • Tcf7l2 protein, mouse
  • Trans-Activators
  • Transcription Factor 4
  • Transcription Factor 7-Like 2 Protein
  • Transcription Factors
  • Luciferases
  • Glutathione Transferase