Exp Mol Med.  2002 Nov;34(5):367-373.

Inhibition of BETA2/NeuroD by Id2

• Ghil SH
• Jeon YJ
• Suh-Kim H

Abstract

Id (Inhibitor of Differentiation) proteins belong to a family of transcriptional modulators that are characterized by a helix loop helix (HLH) region but lack the basic amino acid domain. Id proteins are known to interact with basic helix-loop-helix (bHLH) transcription factors and function as their negative regulators. The negative role of Id proteins has been well demonstrated in muscle development and some in neuronal cells. In this study, we investigated the effect of Id on the function of BETA2/NeuroD, a bHLH transcription factor responsible for neuron and endocrine cell specific gene expression. cDNAs of several Id isoforms were isolated by yeast two-hybrid system using the bHLH domain of E47, a ubiquitous bHLH partner as a bait. Id proteins expressed in COS M6 cells, were found in both cytosolic and nuclear fractions. Electrophoretic mobility shift assay showed that coexpression of Id2 proteins inhibited BETA2/ NeuroD binding to its target sequence, E-box. Id2 inhibited E-box mediated gene expression in a dose dependent manner in BETA2/NeuroD expressing HIT cells. Id coexpressed with BETA2/NeuroD in HeLa cells, inhibited the stimulatory activity of BETA2/NeuroD. These results suggest that Id proteins may negatively regulate tissue specific gene expression induced by BETA2/NeuroD in neuroendocrine cells and the inhibitory role of Id proteins during differentiation may be conserved in various tissues.

Keyword

bHLH; id; BETA2/NeuroD; neuronal differentiation

MeSH Terms

Amino Acid Sequence
Animals
Base Sequence
Binding Sites
Cells, Cultured
DNA-Binding Proteins/genetics/*metabolism
E-Box Elements
Gene Expression Regulation/physiology
Helix-Loop-Helix Motifs
Human
Islets of Langerhans/cytology/metabolism
Mice
Molecular Sequence Data
Nerve Tissue Proteins/genetics/*metabolism
Neurons/cytology/metabolism
Organ Specificity
Transcription Factors/genetics/*metabolism
Two-Hybrid System Techniques