ZFIN ID: ZDB-PUB-030902-5
Novel regulatory sequence -82/-62 functions as a key element to drive the somite-specificity of zebrafish myf-5
Chen, Y.-H., Lee, H.-C., Liu, C.-F., Lin, C.-Y., and Tsai, H.-J.
Date: 2003
Source: Developmental dynamics : an official publication of the American Association of Anatomists   228(1): 41-50 (Journal)
Registered Authors: Chen, Yau-Hung, Tsai, Huai-Jen
Keywords: none
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Cytomegalovirus/genetics
  • DNA-Binding Proteins*
  • Gene Expression Regulation, Developmental
  • Genes, Regulator
  • Genes, Reporter
  • Green Fluorescent Proteins
  • Luminescent Proteins/metabolism
  • Models, Biological
  • Muscle Proteins/chemistry
  • Muscle Proteins/metabolism*
  • Muscle, Skeletal/growth & development
  • Mutagenesis, Site-Directed
  • Myogenic Regulatory Factor 5
  • Promoter Regions, Genetic*
  • Recombinant Fusion Proteins/metabolism
  • Sequence Deletion
  • Somites/metabolism*
  • Time Factors
  • Trans-Activators*
  • Transcription Factors/chemistry
  • Transcription Factors/metabolism*
  • Zebrafish/embryology*
PubMed: 12950078 Full text @ Dev. Dyn.
ABSTRACT
Myf-5, a transcription factor that controls muscle differentiation, is expressed in somites during early embryogenesis. However, gene regulation of myf-5 is poorly understood and detailed functional analysis of the regulatory cis-elements is needed. In zebrafish, the myf-5 upstream sequence from -82 to -62 (-82/-62) was fused with a basal promoter and transferred to fertilized zebrafish eggs. The -82/-62 cassette drove green fluorescent protein (GFP) reporter gene expression specifically in the somites. Moreover, GFP signals were detected exclusively in the somites of 28-hpf embryos derived from eggs injected with pCMV-5x(-82/-62), which contained five copies of the -82/-62 cassette inserted within cytomegalovirus promoter/enhancer. Thus, the -82/-62 cassette, conserved in mouse myf-5, functions to drive somite-specific expression and to repress nonspecific expression during the early development of zebrafish embryos. Mutated sequence analysis of -82/-62 cassette showed that the -70/-62 sequence was the key element for controlling myf-5 specificity. The putative CCAAT-like box, located at -66/-62, could not direct somite-specific expression. A DNA-protein complex was specifically formed between the -70/-62 probe and embryonic nuclear extracts. We conclude that the -70/-62 motif is essential for controlling somite-specific expression and the CCAAT-like box is essential for activating gene transcription.
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