|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.
|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|
|PubMed:||12950078 Full text @ Dev. Dyn.|
Chen, Y.-H., Lee, H.-C., Liu, C.-F., Lin, C.-Y., and Tsai, H.-J. (2003) Novel regulatory sequence -82/-62 functions as a key element to drive the somite-specificity of zebrafish myf-5. Developmental dynamics : an official publication of the American Association of Anatomists. 228(1):41-50.
ABSTRACTMyf-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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