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Wnt antagonist, secreted frizzled-related protein 1, is involved in prenatal skeletal muscle development and is a target of miRNA-1/206 in pigs.

Yang Y, Sun W, Wang R, Lei C, Zhou R, Tang Z, Li K - BMC Mol. Biol. (2015)

Bottom Line: The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression.Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R. China. yangyalan1988@126.com.

ABSTRACT

Background: The Wnt signaling pathway is involved in the control of cell proliferation and differentiation during skeletal muscle development. Secreted frizzled-related proteins (SFRPs), such as SFRP1, function as inhibitors of Wnt signaling. MicroRNA-1/206(miRNA-1/206) is specifically expressed in skeletal muscle and play a critical role in myogenesis. The miRNA-mRNA profiles and bioinformatics study suggested that the SFRP1 gene was potentially regulated by miRNA-1/206 during porcine skeletal muscle development.

Methods: To understand the function of SFRP1 and miRNA-1/206 in swine myogenesis, we first predicted the targets of miRNA-1/206 with the TargetScan and PicTar programs, and analyzed the molecular characterization of the porcine SFRP1 gene. We performed a temporal-spatial expression analysis of SFRP1 mRNA and miRNA-206 in Tongcheng pigs (a Chinese indigenous breed) by quantitative real-time polymerase chain reaction, and conducted the co-expression analyses of SFRP1 and miRNA-1/206. Subsequently, the interaction between SFRP1 and miRNA-1/206 was validated via dual luciferase and Western blot assays.

Results: The bioinformatics analysis predicted SFRP1 to be a target of miRNA-1/206. The expression level of the SFRP1 was highly varied across numerous pig tissues and it was down-regulated during porcine skeletal muscle development. The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression. A significant negative correlation was observed between the expression of miR-1/206 and SFRP1 during porcine skeletal muscle development (p <0.05). Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.

Conclusions: Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development. These findings increase understanding of the biological functions and the regulation of the SFRP1 gene in mammals.

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Comparison of porcineSFRP1sequence (GenBank accession ID: XP_003359916.2) with those of human (GenBank accession ID: NP_003003.3), mouse (GenBank accession ID: NP_038862.2) and rat (GenBank accession no. NP_001263641.1) sequence. Shading shows identical and similar amino acid residues among the four species. Common structural domains are indicated by boxes including a Frizzled domain and a Netrin C-terminal domain.
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Fig2: Comparison of porcineSFRP1sequence (GenBank accession ID: XP_003359916.2) with those of human (GenBank accession ID: NP_003003.3), mouse (GenBank accession ID: NP_038862.2) and rat (GenBank accession no. NP_001263641.1) sequence. Shading shows identical and similar amino acid residues among the four species. Common structural domains are indicated by boxes including a Frizzled domain and a Netrin C-terminal domain.

Mentions: The SFRP1 protein sequences (Accession ID: XM_003359868.3) were retrieved from the GenBank database. First, the amino acid sequence of porcine SFRP1 was compared with those of the human, mouse and rat, and the alignment results showed that the protein sequence of the pig had a six amino acid depletion. The porcine SFRP1 polypeptide exhibited 96.09%, 94.26% and 93.31% similarity with the human, mouse and rat homologs, respectively (Figure 2). Based on the phylogenetic tree analysis, the protein sequence of the pig was closely related to that of cow, while the rat and mouse formed another closely related group, and these four species formed a group with the humans (Figure 3A). SMART software was used to predict SFRP1 protein domains. The precursor protein of SFRP1 contained a Frizzled domain and a Netrin C-terminal domain(Figure 3B). The sites of SFRP1 protein localization had a 44.4% possibility of being extracellular, a 22.2% possibility of being cytoplasmic and a 22.2% possibility of being in the endoplasmic reticulum. Additionally, the theoretical isoelectric point (pI) and molecular weight (Mw) of SFRP1 were 9.05 and 34.79 KDa, respectively.Figure 2


Wnt antagonist, secreted frizzled-related protein 1, is involved in prenatal skeletal muscle development and is a target of miRNA-1/206 in pigs.

Yang Y, Sun W, Wang R, Lei C, Zhou R, Tang Z, Li K - BMC Mol. Biol. (2015)

Comparison of porcineSFRP1sequence (GenBank accession ID: XP_003359916.2) with those of human (GenBank accession ID: NP_003003.3), mouse (GenBank accession ID: NP_038862.2) and rat (GenBank accession no. NP_001263641.1) sequence. Shading shows identical and similar amino acid residues among the four species. Common structural domains are indicated by boxes including a Frizzled domain and a Netrin C-terminal domain.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359577&req=5

Fig2: Comparison of porcineSFRP1sequence (GenBank accession ID: XP_003359916.2) with those of human (GenBank accession ID: NP_003003.3), mouse (GenBank accession ID: NP_038862.2) and rat (GenBank accession no. NP_001263641.1) sequence. Shading shows identical and similar amino acid residues among the four species. Common structural domains are indicated by boxes including a Frizzled domain and a Netrin C-terminal domain.
Mentions: The SFRP1 protein sequences (Accession ID: XM_003359868.3) were retrieved from the GenBank database. First, the amino acid sequence of porcine SFRP1 was compared with those of the human, mouse and rat, and the alignment results showed that the protein sequence of the pig had a six amino acid depletion. The porcine SFRP1 polypeptide exhibited 96.09%, 94.26% and 93.31% similarity with the human, mouse and rat homologs, respectively (Figure 2). Based on the phylogenetic tree analysis, the protein sequence of the pig was closely related to that of cow, while the rat and mouse formed another closely related group, and these four species formed a group with the humans (Figure 3A). SMART software was used to predict SFRP1 protein domains. The precursor protein of SFRP1 contained a Frizzled domain and a Netrin C-terminal domain(Figure 3B). The sites of SFRP1 protein localization had a 44.4% possibility of being extracellular, a 22.2% possibility of being cytoplasmic and a 22.2% possibility of being in the endoplasmic reticulum. Additionally, the theoretical isoelectric point (pI) and molecular weight (Mw) of SFRP1 were 9.05 and 34.79 KDa, respectively.Figure 2

Bottom Line: The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression.Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R. China. yangyalan1988@126.com.

ABSTRACT

Background: The Wnt signaling pathway is involved in the control of cell proliferation and differentiation during skeletal muscle development. Secreted frizzled-related proteins (SFRPs), such as SFRP1, function as inhibitors of Wnt signaling. MicroRNA-1/206(miRNA-1/206) is specifically expressed in skeletal muscle and play a critical role in myogenesis. The miRNA-mRNA profiles and bioinformatics study suggested that the SFRP1 gene was potentially regulated by miRNA-1/206 during porcine skeletal muscle development.

Methods: To understand the function of SFRP1 and miRNA-1/206 in swine myogenesis, we first predicted the targets of miRNA-1/206 with the TargetScan and PicTar programs, and analyzed the molecular characterization of the porcine SFRP1 gene. We performed a temporal-spatial expression analysis of SFRP1 mRNA and miRNA-206 in Tongcheng pigs (a Chinese indigenous breed) by quantitative real-time polymerase chain reaction, and conducted the co-expression analyses of SFRP1 and miRNA-1/206. Subsequently, the interaction between SFRP1 and miRNA-1/206 was validated via dual luciferase and Western blot assays.

Results: The bioinformatics analysis predicted SFRP1 to be a target of miRNA-1/206. The expression level of the SFRP1 was highly varied across numerous pig tissues and it was down-regulated during porcine skeletal muscle development. The expression level of the SFRP1 was significantly higher in the embryonic skeletal compared with postnatal skeletal muscle, whereas miR-206 showed the inverse pattern of expression. A significant negative correlation was observed between the expression of miR-1/206 and SFRP1 during porcine skeletal muscle development (p <0.05). Dual luciferase assay and Western-blot results demonstrated that SFRP1 was a target of miR-1/206 in porcine iliac endothelial cells.

Conclusions: Our results indicate that the SFRP1 gene is regulated by miR-1/206 and potentially affects skeletal muscle development. These findings increase understanding of the biological functions and the regulation of the SFRP1 gene in mammals.

Show MeSH