title: Subcellular localization of different regions of porcine Six1 gene and its expression analysis in C2C12 myoblasts.
abstract: Six1 protein belongs to the Six homeoproteins family, exposing typical domain structure. Although the functions of Six1 have been drawn much attention, the roles of its individual domains are not completely elucidated. Here, we first detected the expression patterns of myogenin, MyoD, Myf5, and Six1 genes using real-time PCR in differentiating C2C12 cells cultured in differentiation medium for 2 or 6 days. The results showed that Six1 gene had the similar expression pattern with myogenin, MyoD, and Myf5 genes, which suggests that it may affect the myogenic differentiation. In order to evaluate the role of distinct domains of Six1 protein in subcellular localization, we constructed a series of truncated vectors tagged with green fluorescent proteins expressing various regions of porcine Six1 protein for subcellular localization analysis. Fluorescence confocal microscopy analysis showed that the different regions of Six1 protein displayed discrete distributions throughout the nucleus and the cytoplasm. The full-length CDS was exclusively localized in the nucleus and the individual HD domain was preferentially distributed to the nucleus both in C2C12 cells and in PK cells. However, the SD domain was diffusely distributed to the cytoplasm and the nucleus, and the localization of SD domain was biased to cytoplasm in C2C12 cells. Taken together, we conclude that the HD domain is important for the nuclear localization of porcine Six1 protein.
authors: Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. wuwangjun2009@hotmail.com
authors: Wangjun
authors: W
authors: Wu
authors: Zhuqing
authors: Z
authors: Ren
authors: Chao
authors: C
authors: Chen
authors: Yang
authors: Y
authors: Liu
authors: Lin
authors: L
authors: Zhang
authors: Zhe
authors: Z
authors: Chao
authors: Bo
authors: B
authors: Zuo
authors: Dequan
authors: D
authors: Xu
authors: Minggang
authors: M
authors: Lei
authors: Yuanzhu
authors: Y
authors: Xiong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism; D051787:Myogenic Regulatory Factor 5 / Q000235:genetics / Q000378:metabolism; D018008:Myogenin / Q000235:genetics / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D034421:Sus scrofa
chemical_list: D018398:Homeodomain Proteins; D017570:MyoD Protein; D051787:Myogenic Regulatory Factor 5; D018008:Myogenin; D010446:Peptide Fragments; D011994:Recombinant Proteins