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Specificity protein 1 regulates gene expression related to fatty acid metabolism in goat mammary epithelial cells.

Zhu J, Sun Y, Luo J, Wu M, Li J, Cao Y - Int J Mol Sci (2015)

Bottom Line: There were no significant expression level differences between the mammary gland tissues collected at lactation and dry-off period.These results were further enhanced by the silencing of SP1.These findings suggest that SP1 may play an important role in fatty acid metabolism.

View Article: PubMed Central - PubMed

Affiliation: Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China. zhujiang4656@hotmail.com.

ABSTRACT
Specificity protein 1 (SP1) is a ubiquitous transcription factor that plays an important role in controlling gene expression. Although important in mediating the function of various hormones, the role of SP1 in regulating milk fat formation remains unknown. To investigate the sequence and expression information, as well as its role in modulating lipid metabolism, we cloned SP1 gene from mammary gland of Xinong Saanen dairy goat. The full-length cDNA of the SP1 gene is 4376 bp including 103 bp of 5'UTR, 2358 bp of ORF (HM_236311) and 1915 bp of 3'UTR, which is predicted to encode a 786 amino acids polypeptide. Phylogenetic tree analysis showed that goat SP1 has the closest relationship with sheep, followed by bovines (bos taurus, odobenus and ceratotherium), pig, primates (pongo, gorilla, macaca and papio) and murine (rattus and mus), while the furthest relationship was with canis and otolemur. Expression was predominant in the lungs, small intestine, muscle, spleen, mammary gland and subcutaneous fat. There were no significant expression level differences between the mammary gland tissues collected at lactation and dry-off period. Overexpression of SP1 in goat mammary epithelial cells (GMECs) led to higher mRNA expression level of peroxisome proliferator-activated receptor-γ (PPARγ) and lower liver X receptor α (LXRα) mRNA level, both of which were crucial in regulating fatty acid metabolism, and correspondingly altered the expression of their downstream genes in GMECs. These results were further enhanced by the silencing of SP1. These findings suggest that SP1 may play an important role in fatty acid metabolism.

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Effect of expression alteration of SP1 on the expression of genes relative to lipid metabolism. (A) Ad-SP1 overexpressed the expression level of SP1 protein; (B) Ad-SP1 overexpressed the mRNA expression level of the SP1 gene; (C) Overexpression of SP1 altered the mRNA expression level of genes related to lipid metabolism in goat mammary epithelial cells (GMECs); (D) siRNA decreased the protein expression of SP1; (E) siRNA decreased the mRNA expression of SP1; and (F) The silencing of SP1 affected the mRNA expression of genes associated with lipid metabolism. Columns, average of 3 repeats; bars, SD; *, p < 0.05
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ijms-16-01806-f004: Effect of expression alteration of SP1 on the expression of genes relative to lipid metabolism. (A) Ad-SP1 overexpressed the expression level of SP1 protein; (B) Ad-SP1 overexpressed the mRNA expression level of the SP1 gene; (C) Overexpression of SP1 altered the mRNA expression level of genes related to lipid metabolism in goat mammary epithelial cells (GMECs); (D) siRNA decreased the protein expression of SP1; (E) siRNA decreased the mRNA expression of SP1; and (F) The silencing of SP1 affected the mRNA expression of genes associated with lipid metabolism. Columns, average of 3 repeats; bars, SD; *, p < 0.05

Mentions: After treatment with adenovirus of Ad-SP1 for 72 h, both the protein (Figure 4A) and mRNA expression (Figure 4B) of SP1 were significantly increased. Accompanying the overexpression of the SP1 gene, an obvious increase was observed in PPARγ expression, while LXRα was decreased dramatically by adenovirus treatment (Figure 4C). Corresponding to the result of SP1 overexpression, silencing of SP1 (Figure 4D,E) decreased the expression of PPARγ and increased the expression of LXRα significantly (Figure 4F). However, SREBP1 remained unaffected after adenovirus or siRNA treatments, though a slight decrease was observed (Figure 4C,F).


Specificity protein 1 regulates gene expression related to fatty acid metabolism in goat mammary epithelial cells.

Zhu J, Sun Y, Luo J, Wu M, Li J, Cao Y - Int J Mol Sci (2015)

Effect of expression alteration of SP1 on the expression of genes relative to lipid metabolism. (A) Ad-SP1 overexpressed the expression level of SP1 protein; (B) Ad-SP1 overexpressed the mRNA expression level of the SP1 gene; (C) Overexpression of SP1 altered the mRNA expression level of genes related to lipid metabolism in goat mammary epithelial cells (GMECs); (D) siRNA decreased the protein expression of SP1; (E) siRNA decreased the mRNA expression of SP1; and (F) The silencing of SP1 affected the mRNA expression of genes associated with lipid metabolism. Columns, average of 3 repeats; bars, SD; *, p < 0.05
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307335&req=5

ijms-16-01806-f004: Effect of expression alteration of SP1 on the expression of genes relative to lipid metabolism. (A) Ad-SP1 overexpressed the expression level of SP1 protein; (B) Ad-SP1 overexpressed the mRNA expression level of the SP1 gene; (C) Overexpression of SP1 altered the mRNA expression level of genes related to lipid metabolism in goat mammary epithelial cells (GMECs); (D) siRNA decreased the protein expression of SP1; (E) siRNA decreased the mRNA expression of SP1; and (F) The silencing of SP1 affected the mRNA expression of genes associated with lipid metabolism. Columns, average of 3 repeats; bars, SD; *, p < 0.05
Mentions: After treatment with adenovirus of Ad-SP1 for 72 h, both the protein (Figure 4A) and mRNA expression (Figure 4B) of SP1 were significantly increased. Accompanying the overexpression of the SP1 gene, an obvious increase was observed in PPARγ expression, while LXRα was decreased dramatically by adenovirus treatment (Figure 4C). Corresponding to the result of SP1 overexpression, silencing of SP1 (Figure 4D,E) decreased the expression of PPARγ and increased the expression of LXRα significantly (Figure 4F). However, SREBP1 remained unaffected after adenovirus or siRNA treatments, though a slight decrease was observed (Figure 4C,F).

Bottom Line: There were no significant expression level differences between the mammary gland tissues collected at lactation and dry-off period.These results were further enhanced by the silencing of SP1.These findings suggest that SP1 may play an important role in fatty acid metabolism.

View Article: PubMed Central - PubMed

Affiliation: Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China. zhujiang4656@hotmail.com.

ABSTRACT
Specificity protein 1 (SP1) is a ubiquitous transcription factor that plays an important role in controlling gene expression. Although important in mediating the function of various hormones, the role of SP1 in regulating milk fat formation remains unknown. To investigate the sequence and expression information, as well as its role in modulating lipid metabolism, we cloned SP1 gene from mammary gland of Xinong Saanen dairy goat. The full-length cDNA of the SP1 gene is 4376 bp including 103 bp of 5'UTR, 2358 bp of ORF (HM_236311) and 1915 bp of 3'UTR, which is predicted to encode a 786 amino acids polypeptide. Phylogenetic tree analysis showed that goat SP1 has the closest relationship with sheep, followed by bovines (bos taurus, odobenus and ceratotherium), pig, primates (pongo, gorilla, macaca and papio) and murine (rattus and mus), while the furthest relationship was with canis and otolemur. Expression was predominant in the lungs, small intestine, muscle, spleen, mammary gland and subcutaneous fat. There were no significant expression level differences between the mammary gland tissues collected at lactation and dry-off period. Overexpression of SP1 in goat mammary epithelial cells (GMECs) led to higher mRNA expression level of peroxisome proliferator-activated receptor-γ (PPARγ) and lower liver X receptor α (LXRα) mRNA level, both of which were crucial in regulating fatty acid metabolism, and correspondingly altered the expression of their downstream genes in GMECs. These results were further enhanced by the silencing of SP1. These findings suggest that SP1 may play an important role in fatty acid metabolism.

Show MeSH