Abstract
Myostatin is a negative regulator of myogenesis and has been suggested to be an important factor in the development of muscle wasting during viral infection. The objective of this study was to characterize the main regulatory element of the grouper myostatin promoter and to study changes in promoter activity due to viral stimulation. In vitro and in vivo experiments indicated that the E-box E6 is a positive cis-and trans-regulation motif, and an essential binding site for MyoD. In contrast, the E-box E5 is a dominant negative cis-regulatory. The characteristics of grouper myostatin promoter are similar in regulation of muscle growth to that of other species, but mainly through specific regulatory elements. According to these results, we conducted a study to investigate the effect of viral infection on myostatin promoter activity and its regulation. The nervous necrosis virus (NNV) treatment significantly induced myostatin promoter activity. The present study is the first report describing that specific myostatin motifs regulate promoter activity and response to viral infection.
Highlights
Myostatin (GDF-8) is a member of the transforming growth factor-β (TGF-β) superfamily and acts as a negative regulator of skeletal muscle growth and development [1,2,3,4]
The present study found, using deletion analysis, that E-box E6 was the most important positive cis-element; deletion analysis of E-box E6 resulted in a significant reduction in promoter activity
The results of both deletion and truncation analyses indicated that the E-box E6 region performed a dominant positive effect upon the activation of orange-spotted grouper myostatin promoter
Summary
Myostatin (GDF-8) is a member of the transforming growth factor-β (TGF-β) superfamily and acts as a negative regulator of skeletal muscle growth and development [1,2,3,4]. The characterization and expression of the myostatin gene has been studied in several fish species, such as channel catfish (Ictalurus punctatus), yellow catfish (Pelteobagrus fulvidraco), zebrafish (Danio rerio), gilthead seabream (Sparus aurata), orange-spotted grouper (Epinephelus coioides), Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss), Mozambique tilapia (Oreochromis mossambicus), white bass (Morone chrysops), striped bass (Morone saxatilis) and white perch (Morone americana) [5,6,7,8,9,10,11,12,13,14]. We previously reported that the full orange-spotted grouper myostatin mRNA length is 2776 bp with three exons. Its promoter includes 1974 bp located 5’ upstream of the initiation site of myostatin mRNA, and contains ten E-box motifs. Viral infection upregulates myostatin promoter activity study design, data collection and analysis, decision to publish, or preparation of the manuscript
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