Transcriptional repression by the basic helix-loop-helix protein Dec2: multiple mechanisms through E-box elements.
Dec2, a member of the basic helix-loop-helix (bHLH) superfamily, has been shown to function as a transcriptional repressor and is implicated in cell proliferation and differentiation. In addition, Dec2 transcripts exhibit a striking circadian oscillation in the suprachiasmatic nucleus. To identify the molecular mechanisms by which Dec2 regulates gene expression, we carried out structure-function analyses. Gel retardation and luciferase assays showed that Dec2, as well as its related protein Dec1, preferentially binds to class B E-box elements (CACGTG) as a homodimer and represses the transcription of target genes in a histone deacetylase (HDAC)-dependent manner. Functional studies with the GAL4-DNA binding domain fusion proteins identified the domain responsible for the repression activity of Dec2 in its C-terminal region, which is also necessary to recruit HDAC1. In addition, the basic and HLH domains of Dec2 were required for DNA binding and homodimerization, respectively. In contrast, Dec proteins repressed a MyoD-activated promoter activity of muscle creatine kinase gene through class A E-box in an HDAC1-independent manner. Dec2 formed a heterodimer with MyoD through the basic and HLH domains. Consistent with this, both the basic and HLH domains were required for the ability of Dec2 to inhibit the transcriptional activity of MyoD. These findings indicate that Dec2 employs multiple mechanisms, including DNA-binding and protein-protein interactions, to achieve E-box-dependent transcriptional repressions.
Fujimoto
Katsumi
K
Department of Dental and Medical Biochemistry, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima 734-8553, Japan.
Hamaguchi
Hidenori
H
Hashiba
Takafumi
T
Nakamura
Tadahiro
T
Kawamoto
Takeshi
T
Sato
Fuyuki
F
Noshiro
Mitsuhide
M
Bhawal
Ujjal K
UK
Suardita
Ketut
K
Kato
Yukio
Y
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism / Q000502:physiology; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D052283:Creatine Kinase, MM Form / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D019281:Dimerization; D024721:E-Box Elements; D005786:Gene Expression Regulation; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases / Q000378:metabolism / Q000502:physiology; D018398:Homeodomain Proteins / Q000378:metabolism / Q000502:physiology; D051379:Mice; D017570:MyoD Protein / Q000502:physiology; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D056564:Sirtuin 1; D037761:Sirtuins / Q000378:metabolism / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism* / Q000502:physiology*
D051792:Basic Helix-Loop-Helix Proteins; D052283:Creatine Kinase, MM Form; D004268:DNA-Binding Proteins; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases; D018398:Homeodomain Proteins; D017570:MyoD Protein; D056564:Sirtuin 1; D037761:Sirtuins; D014157:Transcription Factors; C470968:Bhlhb3 protein, mouse; C491539:Bhlhe40 protein, mouse; C057363:MyoD1 myogenic differentiation protein; C470545:Sirt1 protein, mouse; C484389:Hdac1 protein, mouse