Cloning, Expression and Function Analysis of Chicken FBXO38t1

Abstract

F-box proteins are widely present in eukaryotes,and their biological functions are diverse in animals.In this study,the full-length coding sequence of an unnamed gene LOC416151(GenBank accession XM_414482) was cloned from the chicken abdominal adipose tissue by reverse transcriptase(RT)-PCR,and the sequence analysis showed that the acquired sequence(GenBank accession.JX290204) is one of the transcript variants of chicken FBXO38,which was designated as gFBXO38t1.Real-time RT-PCR analysis showed that gFBXO38t1 was widely expressed in various chicken tissues,with a relatively higher expression level in the pancreas,ileum and abdominal fat tissue.In addition,gFBXO38t1 was expressed in all the chicken abdominal fat tissues used in the present study,and at 3 and 4 weeks of age,the gFBXO38t1 expression in lean males was significantly greater than that in fat males(P 0.05) and no significant difference was observed at the other ages(P 0.05).The gFBXO38t1 expression decreased followed by the differentiation of chicken preadipocytes induced by oleate,and gFBXO38t1 expressed more highly in chicken preadipocytes than in mature adipocytes(P 0.05),suggesting that gFBXO38t1 might play a negative role in the chicken adipogenesis.Additionally,The luciferase reporter assay showed that gFBXO38t1 overexpression inhibited the promoter activities of chicken CCAAT/enhancer-binding protein α(C/EBPα),lipoprotein lipase(LPL),fatty acid synthase(FASN) and fatty acid-binding protein 4(FABP4).The combined overexpression of gKLF7 and gFBXO38t1 did not lead to an enhanced ability for gFBXO38t1 to regulate the promoter activities of chicken LPL,FABP4 and FASN,indicating that gFBXO38t1 may regulate promoter activities of these genes through a KLF7-independent manner.The current study provides evidence that the gFBXO38t1 is involved in chicken adipogenesis.