Molecular Cloning and Characterization of DEC2, a New Member of Basic Helix-Loop-Helix Proteins

doi:10.1006/bbrc.2000.4133

Biochemical and Biophysical Research Communications

Volume 280, Issue 1, 12 January 2001, Pages 164-171

https://doi.org/10.1006/bbrc.2000.4133 Get rights and content

Abstract

DEC1 is a basic helix-loop-helix (bHLH) protein related to Drosophila Hairy, Enhancer of split and HES, and involved in the control of proliferation and/or differentiation of chondrocytes, neurons, etc. We report here the identification and characterization of human, mouse and rat DEC2, a novel member of the DEC subfamily. DEC2 had high (97%) and moderate (52%) similarities in the bHLH region and the Orange domain with DEC1, respectively. However, DEC2, but not DEC1, had alanine and glycine-rich regions in the C-terminal half. Unlike Hairy, Enhancer of split and HES, DEC2 lacked the WRPW motif for interaction with the corepressor Groucho. The DEC2 gene was mapped to human chromosome 12p11.23-p12.1, mouse chromosome 6 G2-G3 and rat chromosome 4q43 distal–q4, where the conserved linkage homology has been identified among these species. Unlike DEC1, which was broadly expressed in many tissues, DEC2 showed a more restricted pattern of mRNA expression. The DEC subfamily proteins may play an important role in tissue development.

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Smad3 Suppresses Epithelial Cell Migration and Proliferation via the Clock Gene Dec1, Which Negatively Regulates the Expression of Clock Genes Dec2 and Per1
2019, American Journal of Pathology
Smad3 has circadian expression; however, whether Smad3 affects the expression of clock genes is poorly understood. Here, we investigated the regulatory mechanisms between Smad3 and the clock genes Dec1, Dec2, and Per1. In Smad3 knockout mice, the amplitude of locomotor activity was decreased, and Dec1 expression was decreased in the suprachiasmatic nucleus, liver, kidney, and tongue compared with control mice. Conversely, Dec2 and Per1 expression was increased compared with that of control mice. In Smad3 knockout mice, immunohistochemical staining revealed that Dec1 expression decreased, whereas Dec2 and Per1 expression increased in the endothelial cells of the kidney and liver. In NIH3T3 cells, Smad3 overexpression increased Dec1 expression, but decreased Dec2 and Per1 expression. In a wound-healing experiment that used Smad3 knockout mice, Dec1 expression decreased in the basal cells of squamous epithelium, promoting wound healing of the mucosa. Finally, the migration and proliferation of Smad3 knockdown squamous carcinoma cells was suppressed by Dec1 overexpression but was promoted by Dec2 overexpression. Dec1 overexpression decreased E-cadherin and proliferating cell nuclear antigen expression, whereas these expression levels were increased by Dec2 overexpression. These results suggest Smad3 is relevant to circadian rhythm and regulates cell migration and proliferation through Dec1, Dec2, and Per1 expression.
Differentiated embryonic chondrocytes 1 expression of periodontal ligament tissue and gingival tissue in the patients with chronic periodontitis
2015, Archives of Oral Biology
To evaluate the DEC1 expression of periodontal ligament tissue and gingival tissue in the patients with chronic periodontitis.
20 non-smoking patients with chronic periodontitis and 20 healthy individuals were enrolled. Periodontal ligament tissue and gingival tissue samples from healthy subjects were collected during teeth extraction for orthodontic reason or the third molar extraction. The parallel samples from patients with chronic periodontitis were obtained during periodontal flap operations or teeth extraction as part of periodontal treatment. The DEC1 expression and the alkaline phosphatase (ALP) activity of both the periodontal ligament tissue and gingival tissue were determined by Western blot, Immunohistochemistry and ALP Detection Kit.
The DEC1 expression of periodontal ligament tissue in the patients with chronic periodontitis decreased significantly along with the decreased ALP activity. On the contrary, the DEC1 expression of gingival tissue in the patients with chronic periodontitis increased significantly. Further study found that the DEC1 expression of gingival tissue increased mainly in the suprabasal layer of gingival epithelial cells but decreased in the gingival connective tissue of the patients with chronic periodontitis.
The DEC1 expression decreases in the periodontal ligament tissue which is related to the osteogenic capacity, whereas the DEC1 expression increases in the suprabasal layer of gingival epithelial cells which are involved in immune inflammatory response in the patients with chronic periodontitis. The findings provide a new target to explore the pathology and the therapy of periodontitis.
The basic helix-loop-helix (bHLH) transcription factor DEC2 negatively regulates Twist1 through an E-box element
2014, Biochemical and Biophysical Research Communications
Differentiated embryo chondrocyte 2 (DEC2/Sharp-1/Bhlhe41), a basic helix-loop-helix (bHLH) transcription factor, has been shown to regulate the transcription of target genes by binding to their E-box elements. We identified a possible DEC2-response element (consensus E-box: CACGTG) in the promoter region of Twist1. Forced expression of DEC2 significantly repressed Twist1 promoter activity under normoxia and under hypoxia as assessed by a luciferase reporter assay. In addition, over-expression of DEC2 repressed Twist1 mRNA expression assessed by quantitative real-time PCR. Site-directed mutagenesis studies showed that mutagenesis of the consensus E-box sequence eliminated the ability of DEC2 to reduce the Twist1 promoter activity. Chromatin immunoprecipitation (ChIP) assays confirmed that the DEC2-mediated repression is primarily achieved by binding to the E-box in the Twist1 promoter. Knockdown of DEC2 by siRNA significantly attenuated the repression of Twist1 expression. DEC2 and Twist1 exhibit inversed protein expression patterns during development of mouse tongue embryo tissue. Given the fact that DEC2 protein is emerging as an important regulator in a vast array of cellular events, including cell differentiation, maturation of lymphocytes and the molecular clock, our study elucidates an important mechanism by which DEC2 regulates cellular function by modulating the expression of Twist1.
DEC1/STRA13/SHARP2 and DEC2/SHARP1 coordinate physiological processes, including circadian rhythms in response to environmental stimuli
2014, Current Topics in Developmental Biology
Citation Excerpt :
Thus, DEC1 (differentiated embryonic chondrocyte 1) is also referred to as BHLHe40 (Shen et al., 1997), Stimulated with retinoic acid 13 (Stra13) (Boudjelal et al., 1997), E47-interacting protein 1 (Eip1) (Dear et al., 1997), Enhancer-of-Split (E(spl)) and Hairy-related protein-2 (Sharp-2) (Rossner, Dorr, Gass, Schwab, & Nave, 1997), cytokine response gene 8 (CR8) (Beadling, Cereseto, Fan, Naramura, & Smith, 2001), or Clast5 (Seimiya et al., 2002). DEC2 is also called BHLHe41 or Sharp-1 (Rossner et al., 1997; Fujimoto et al., 2001). DEC1 and DEC2 may play a role in the adaptation to environmental signals, such as day–night cycle, availability of nutrients, oxygen concentration, infection, and ionizing radiation.
Daily physiological and behavioral rhythms are regulated by endogenous circadian molecular clocks. Clock proteins DEC1 (BHLHe40) and DEC2 (BHLHe41) belong to the basic helix–loop–helix protein superfamily, which contains other clock proteins CLOCK and BMAL1. DEC1 and DEC2 are induced by CLOCK:BMAL1 heterodimer via the CACGTG E-box in the promoter and, thereafter, suppress their own expression by competing with CLOCK:BMAL1 for the DNA binding. This negative feedback DEC loop together with the PER loop involving PER and CRY, the other negative clock regulators, maintains the circadian rhythm of Dec1 and Dec2 expression. DEC1 is induced by light pulse and adjusts the circadian phase of the central clock in the suprachiasmatic nucleus, whereas DEC1 upregulation by TGF-β resets the circadian phase of the peripheral clocks in tissues. Furthermore, DEC1 and DEC2 modulate the clock output signals to control circadian rhythms in behavior and metabolism. In addition to the functions in the clocks, DEC1 and DEC2 are involved in hypoxia responses, immunological reactions, and carcinogenesis. These DEC actions are mediated by the direct binding to the E-box elements in target genes or by protein–protein interactions with transcription factors such as HIF-1α, RXRα, MyoD, and STAT. Notably, numerous growth factors, hormones, and cytokines, along with ionizing radiation and DNA-damaging agents, induce Dec1 and/or Dec2 in a tissue-specific manner. These findings suggest that DEC1 and DEC2 play a critical role in animal adaptation to various environmental stimuli.
Stra13 and Sharp-1, the non-grouchy regulators of development and disease
2014, Current Topics in Developmental Biology
Citation Excerpt :
A highly related gene Sharp-1 was also isolated from rat brain (Rossner et al., 1997). Subsequent studies led to the cloning of its human and mouse counterparts named Dec2 and Sharp-1 (Azmi & Taneja, 2002; Fujimoto et al., 2001). Orthologues for Stra13/Dec1/Sharp-2 and Sharp-1/Dec2 have also been identified in zebrafish (Chen, Zhou, Xu, Xu, & Xue, 2010; Yao et al., 2006) as well as in Drosophila melanogaster, which contains a single gene named clockwork orange (cwo) (Kadener, Stoleru, McDonald, Nawathean, & Rosbash, 2007).
Stra13 and Sharp-1 are transcriptional repressors that share domain structure similarity with members of the basic helix-loop-helix-Orange subfamily. In contrast to other members that include Hes and Hey proteins, transcriptional repression mediated by Stra13 and Sharp-1 does not involve recruitment of the corepressor Groucho. Both proteins undergo sumoylation at evolutionarily conserved sites, and this posttranslational modification serves as a platform for association with chromatin-modifying enzymes including histone deacetylases and histone methyltransferases. In addition to being widely expressed during embryonic development and in adult tissues, the expression of both genes is induced by a number of stimuli. Loss-of-function and gain-of-function studies have demonstrated their function in cellular differentiation and regeneration, in regulation of circadian rhythms, immune homeostasis, and metabolism. Given their diverse physiological functions in several tissues, it is not surprising that deregulated expression of Stra13 and Sharp-1 is apparent in human pathologies. Here, we review our current understanding of their cellular functions that suggest a requirement in maintenance of tissue homeostasis.
Role of differentiated embryo-chondrocyte expressed gene 2 in immunity
2024, Frontiers in Immunology

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^☆: The nucleotide sequences of human and mouse DEC2 have been submitted to GenBank under Accession Nos. AB044088 and AB044090, respectively.

¹: To whom correspondence and reprint requests should be addressed. Fax: +81-82-257-5629. E-mail: [email protected].

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Regular ArticleMolecular Cloning and Characterization of DEC2, a New Member of Basic Helix-Loop-Helix Proteins☆

Abstract

Biochim. Biophys. Acta

Biochem. Biophys. Res. Commun.

Mol. Cell. Neurosci.

Biochem. Biophys. Res. Commun.

Cell

Anal. Biochem.

Cell

Biochem. Biophys. Res. Commun.

Mech. Dev.

Dev. Biol.

Genomics

Cancer Genet. Cytogenet.

Overexpression of Stra13, a novel retinoic acid-inducible gene of the basic helix-loop-helix family, inhibits mesodermal and promotes neuronal differentiation of P19 cells

Genes Dev.

Stra13 expression is associated with growth arrest and represses transcription through histone deacetylase (HDAC)-dependent and HDAC-independent mechanisms

Proc. Natl. Acad. Sci. USA

The function of hairy-related bHLH repressor proteins in cell fate decisions

BioEssays

R-banding and nonisotopic in situ hybridization: Precise localization of the human type II collagen gene (COL2A1)

Hum. Genet.

Regular Article
Molecular Cloning and Characterization of DEC2, a New Member of Basic Helix-Loop-Helix Proteins☆