title: Genomic cloning and promoter functional analysis of myostatin-2 in shi drum, Umbrina cirrosa: conservation of muscle-specific promoter activity.
abstract: Myostatin (MSTN) is a member of the transforming growth factor-ß superfamily, known as a negative regulator of skeletal muscle development and growth in mammals. In contrast to mammals, fish possess at least two paralogs of MSTN: MSTN-1 and MSTN-2. Here we describe the cloning and sequence analysis of spliced and precursor (unspliced) transcripts as well as the 5' flanking region of MSTN-2 from the marine fish Umbrina cirrosa (ucMSTN-2). In silico analysis revealed numerous putative cis regulatory elements including several E-boxes known as binding sites to myogenic transcription factors. Transient transfection experiments using non-muscle and muscle cell lines showed high transcriptional activity in muscle cells and in differentiated neural cells, in accordance with our previous findings in MSTN-2 promoter from Sparus aurata. Comparative informatics analysis of MSTN-2 from several fish species revealed high conservation of the predicted amino acid sequence as well as the gene structure (exon length) although intron length varied between species. The proximal promoter of MSTN-2 gene was found to be conserved among Perciforms. In conclusion, this study reinforces our conclusion that MSTN-2 promoter is a very strong promoter, especially in muscle cells. In addition, we show that the MSTN-2 gene structure is highly conserved among fishes as is the predicted amino acid sequence of the peptide.
authors.affiliation: Department of Marine Biology & Biotechnology, National Institute of Oceanography, Israel Oceanographic and Limnological Research, Tel-Shikmona, P.O.B 8030, Haifa 31080, Israel.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Nadjar-Boger
authors.forename: Lisa
authors.initials: L
authors.lastname: Maccatrozzo
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Radaelli
authors.forename: Bruria
authors.initials: B
authors.lastname: Funkenstein
mesh_terms: D024506:5' Flanking Region / Q000235:genetics; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D029941:Fish Proteins / Q000145:classification / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D055435:Myostatin / Q000145:classification / Q000235:genetics*; D010473:Perciformes / Q000235:genetics*; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D029941:Fish Proteins; D055435:Myostatin; D020033:Protein Isoforms; D011498:Protein Precursors