Cloning, identification and functional analysis of the goat transcription factor <i>c-fos</i>.

Tingting HU; Yong Wang; Dingshuang Chen; Chengsi Gong; Yanyan LI; Yan Xiong; Jianmei Wang; Zhixiong LI; Yaqiu Lin

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Cloning, identification and functional analysis of the goat transcription factor c-fos.

Author: Tingting HU ¹ ; Yong WANG ¹ ; Dingshuang CHEN ¹ ; Chengsi GONG ¹ ; Yanyan LI ¹ ; Yan XIONG ¹ ; Jianmei WANG ¹ ; Zhixiong LI ¹ ; Yaqiu LIN ¹
Author Information

1. College of Animal Science and Veterinary Medicine, Southwest Minzu University, Chengdu 610041, Sichuan, China.
Publication Type:Journal Article
Keywords: c-fos; cell differentiation; goat; subcutaneous fat; transcription factor
MeSH: Animals; Goats/genetics*; Cell Differentiation/genetics*; Adipogenesis/genetics*; Gene Expression Regulation; Proteins/genetics*; Cloning, Molecular
From: Chinese Journal of Biotechnology 2023;39(4):1684-1695
CountryChina
Language:Chinese
Abstract: C-fos is a transcription factor that plays an important role in cell proliferation, differentiation and tumor formation. The aim of this study was to clone the goat c-fos gene, clarify its biological characteristics, and further reveal its regulatory role in the differentiation of goat subcutaneous adipocytes. We cloned the c-fos gene from subcutaneous adipose tissue of Jianzhou big-eared goats by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed its biological characteristics. Using real-time quantitative PCR (qPCR), we detected the expression of c-fos gene in the heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi and subcutaneous adipocytes of goat upon induced differentiation for 0 h to 120 h. The goat overexpression vector pEGFP-c-fos was constructed and transfected into the subcutaneous preadipocytes for induced differentiation. The morphological changes of lipid droplet accumulation were observed by oil red O staining and bodipy staining. Furthermore, qPCR was used to test the relative mRNA level of the c-fos overexpression on adipogenic differentiation marker genes. The results showed that the cloned goat c-fos gene was 1 477 bp in length, in which the coding sequence was 1 143 bp, encoding a protein of 380 amino acids. Protein structure analysis showed that goat FOS protein has a basic leucine zipper structure, and subcellular localization prediction suggested that it was mainly distributed in the nucleus. The relative expression level of c-fos was higher in the subcutaneous adipose tissue of goats (P < 0.05), and the expression level of c-fos was significantly increased upon induced differentiation of subcutaneous preadipocyte for 48 h (P < 0.01). Overexpression of c-fos significantly inhibited the lipid droplets formation in goat subcutaneous adipocytes, significantly decreased the relative expression levels of the AP2 and C/EBPβ lipogenic marker genes (P < 0.01). Moreover, AP2 and C/EBPβ promoter are predicted to have multiple binding sites. In conclusion, the results indicated that c-fos gene was a negative regulatory factor of subcutaneous adipocyte differentiation in goats, and it might regulate the expression of AP2 and C/EBPβ gene expression.