title: Transcription Factor Sp1 Promotes the Expression of Porcine ROCK1 Gene.
abstract: Rho-associated, coiled-coil containing protein kinase 1 (ROCK1) gene plays a crucial role in maintaining genomic stability, tumorigenesis and myogenesis. However, little is known about the regulatory elements governing the transcription of porcine ROCK1 gene. In the current study, the transcription start site (TSS) was identified by 5'-RACE, and was found to differ from the predicted one. The region in ROCK1 promoter which is critical for promoter activity was investigated via progressive deletions. Site-directed mutagenesis indicated that the region from -604 to -554 bp contains responsive elements for Sp1. Subsequent experiments showed that ROCK1 promoter activity is enhanced by Sp1 in a dose-dependent manner, whereas treatment with specific siRNA repressed ROCK1 promoter activity. Electrophoretic mobility shift assay (EMSA), DNA pull down and chromatin immunoprecipitation (ChIP) assays revealed Sp1 can bind to this region. qRT-PCR and Western blotting research followed by overexpression or inhibition of Sp1 indicate that Sp1 can affect endogenous ROCK1 expression at both mRNA and protein levels. Overexpression of Sp1 can promote the expression of myogenic differentiation 1(MyoD), myogenin (MyoG), myosin heavy chain (MyHC). Taken together, we conclude that Sp1 positively regulates ROCK1 transcription by directly binding to the ROCK1 promoter region (from -604 to -532 bp) and may affect the process of myogenesis.
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. zhangruirui807870@163.com.
authors.forename: Ruirui
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. xiaotingfeng1984@163.com.|College of Life Science and Technology, Wuhan Bioengineering Institute, Wuhan 430070, China. xiaotingfeng1984@163.com.
authors.forename: Xiaoting
authors.initials: X
authors.lastname: Feng
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. zmsyx.2581@163.com.
authors.forename: Mengsi
authors.initials: M
authors.lastname: Zhan
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. huangcong19880426@126.com.
authors.forename: Cong
authors.initials: C
authors.lastname: Huang
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. kunchen1989@163.com.
authors.forename: Kun
authors.initials: K
authors.lastname: Chen
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. wlifetxy@sina.com.
authors.forename: Xiaoyin
authors.initials: X
authors.lastname: Tang
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. 13163228175@163.com.
authors.forename: Tingting
authors.initials: T
authors.lastname: Kang
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. xiongyzhu@sina.com.
authors.forename: Yuanzhu
authors.initials: Y
authors.lastname: Xiong
authors.affiliation: Key Laboratory of Swine Genetics and Breeding of Agricultural Ministry &amp; Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. leimg@mail.hzau.edu.cn.
authors.forename: Minggang
authors.initials: M
authors.lastname: Lei
mesh_terms: D000818:Animals; D002460:Cell Line; D051379:Mice; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D020218:Response Elements; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D054460:rho-Associated Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D016329:Sp1 Transcription Factor; D054460:rho-Associated Kinases; D018995:Myosin Heavy Chains
keywords: ROCK1; Sp1; regulation; transcription factor