Abstract
Proliferation and differentiation of preadipocyte are essential for the formation of fat tissues. However, the genes that regulate the early stage of preadipocyte differentiation in chicken have remained elusive. Here we identify a novel spliced variant of the DNA methyltransferase Dnmt3a gene, named Dnmt3a3, that controls early preadipocyte differentiation. Dnmt3a3 expression is increased at the onset of preadipocyte differentiation and remains elevated during differentiation. Overexpression of Dnmt3a3 in preadipocytes markedly inhibits proliferation and cell-cycle progression, and this is accompanied by inhibition of the mRNA and protein level of cell-cycle control genes, such as p21 and p27. In addition, forced expression of Dnmt3a3 in differentiating preadipocytes represses early preadipocyte differentiation, and this was found to be accompanied by inhibition of the mRNA expression levels of early preadipocyte differentiation markers, such as GATA2, GATA3, C/EBPα, C/EBPβ, AP2, and PPARγ, or the protein levels of GATA3, C/EBPβ, and PPARγ. Taken together, these data demonstrate the participation of Dnmt3a3 in the proliferation and differentiation process of chicken primary preadipocyte cells.
Highlights
Preadipocyte proliferation and differentiation play an essential role in forming adipose tissue
We report the identification of a novel spliced variant of the DNA methyltransferase 3a (Dnmt3a) gene, named Dnmt3a3, that controls early preadipocyte differentiation
The mRNA level of p27, another cell cycle regulator, peaked by day 1 of differentiation and decreased thereafter throughout the course of differentiation, whereas its protein level (22-kDa) gradually elevated throughout the course of differentiation (Figures 3C–E). These results indicate that the mRNA level of Dnmt3a3 was increased at the onset of preadipocyte differentiation and remained elevated throughout the time-course of differentiation
Summary
Preadipocyte proliferation and differentiation play an essential role in forming adipose tissue. Studying the molecular genetics and cellular roles underlying adipose tissue development would improve our understanding of developmental and metabolic syndromes. The central machines that drive cell cycle progression are the cyclin-dependent kinases (CDKs) belonging to the CIP/Kip family (Morrison and Farmer, 1999), such as p21 ( known as Cdkn1a and Cip1) and p27 ( known as Cdkn1b and kip). The central machines that drive cell cycle progression are the cyclin-dependent kinases (CDKs) belonging to the CIP/Kip family (Morrison and Farmer, 1999), such as p21 ( known as Cdkn1a and Cip1) and p27 ( known as Cdkn1b and kip1) These are able to phosphorylate key substrates to promote DNA synthesis and mitotic progression
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