Structural Characterization and Functional Analysis of the Follistatin Promoter of Larimichthys crocea.
Follistatin is a secreted glycoprotein, which involved in numerous physiological activities as an antagonist of the transforming growth factor-β superfamily. However, little is known about the regulation mechanism of follistatin in fish. In this study we cloned and analyzed part of the 5' flanking region of the follistatin gene in Larimichthys crocea. Sequence analysis revealed several putative binding sites for transcription factors including activator protein 1 (AP1), myogenic differentiation factor (MyoD), stimulating protein 1 (SP1), and sex determining gene on the mammalian Y chromosome (SRY) in the cloned fragment. Transcriptional activities of two fragments (485 and 261 bp) truncated from follistatin upstream region were examined in vitro, using transient transfection in Ctenopharyngodon idella kidney (CIK) and Rattus norvegicus skeletal muscle myoblast (L6) cells. The result showed that the promoter activity correlated positively with the length of truncated fragments in both CIK and L6 cells. To study the regulation of follistatin expression in L. crocea, we cloned MyoD and SRY-box 8 (Sox8) genes and examined their action on the follistatin promoter by co-transfection in CIK and L6 cells. The results showed MyoD and Sox8 could suppress the activities of follistatin promoter at different levels in CIK and L6 cells.
1 College of Marine Sciences, Ningbo University , Ningbo, People's Republic of China .|2 Collaborative Innovation Center for Zhejiang Marine High-Efficiency and Healthy Aquaculture, Ningbo University , Ningbo, People's Republic of China .
Huijing
H
Yang
1 College of Marine Sciences, Ningbo University , Ningbo, People's Republic of China .
Zhenhua
Z
Zhang
1 College of Marine Sciences, Ningbo University , Ningbo, People's Republic of China .|2 Collaborative Innovation Center for Zhejiang Marine High-Efficiency and Healthy Aquaculture, Ningbo University , Ningbo, People's Republic of China .
Liangyi
L
Xue
D024506:5' Flanking Region; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D038681:Follistatin / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D010473:Perciformes / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D029941:Fish Proteins; D038681:Follistatin; D007136:Immunoglobulins; D017570:MyoD Protein; D011994:Recombinant Proteins; D055748:SOXB1 Transcription Factors; C090698:SP1 antigen; D018808:Transcription Factor AP-1