Does not hUTP14a promoter form a regulation feedback loop with P53?  

作　　者：Jingyi Zhang Yafei Guo Xiaojuan Du Baocai Xing 

机构地区：[1]Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Hepatic, Biliary & Pancreatic Surgery I,Peking University Cancer Hospital & Institute [2]Department of Cell Biology, Peking University Health Science Center

出　　处：《Chinese Journal of Cancer Research》2014年第2期159-165,共7页中国癌症研究（英文版）

基　　金：supported by grants from Beijing Municipal Natural Science Foundation (Grant No. 7122032);the National Natural Science Foundation of China (Grant No. 81071672);the National Basic Research Program of China (973 program) (Grant No. 2010CB529303);National High Technology Research and Development Program of China (863 Program, Grant No. 2008AA02Z131)

摘　　要：Objective： We previously found that hUTP14a binds P53 and promotes P53 degradation. However, if hUTP14a is a downstream gene of P53 remains to be determined. This study aimed to identify the promoter of h UTP14a and investigate if h UTP14a is regulated by P53. Methods： The hUTPI4a promoter region was cloned into pGL3-Basic-luciferase reporter plasmid to get pGL3-hUTP14a-luc. The reporter plasmid was transfected into 293T cells and luciferase activity was evaluated by the Dual-Luciferase Reporter Assay System. Putative transcription factors were identified through searching Matlnspector Professional and Algorismica i Genetica databases. Either pGL3-hUTPI4aluc or p21 promoter reporter plasmid was co-transfected with increasing dose of p53 plasmid, and luciferase activity was evaluated. A series of deletion constructs of pGL3-hUTP14a-luc were constructed and minimal promoter region of hUTP14a was determined. Differences of the lnciferase activities between different groups were assessed by statistical analysis. Results： The hUTP14a gene promoter reporter construct was correctly cloned and was demonstrated to possess promoter activity. The transcription of hUTP14a was not regulated by P53. The minimal promoter region of h UTP14a gene is located between -203 to -100 of the transcription initiation site. Conclusion： Unlike other P53-interacting proteins such as MDM2, Pirh2 and Cop I which promote P53 degradation and whose transcriptions are regulated by P53, does not hUTP14a transcription form a regulation feedback loop with P 53.Objective： We previously found that hUTP14a binds P53 and promotes P53 degradation. However, if hUTP14a is a downstream gene of P53 remains to be determined. This study aimed to identify the promoter of h UTP14a and investigate if h UTP14a is regulated by P53. Methods： The hUTPI4a promoter region was cloned into pGL3-Basic-luciferase reporter plasmid to get pGL3-hUTP14a-luc. The reporter plasmid was transfected into 293T cells and luciferase activity was evaluated by the Dual-Luciferase Reporter Assay System. Putative transcription factors were identified through searching Matlnspector Professional and Algorismica i Genetica databases. Either pGL3-hUTPI4aluc or p21 promoter reporter plasmid was co-transfected with increasing dose of p53 plasmid, and luciferase activity was evaluated. A series of deletion constructs of pGL3-hUTP14a-luc were constructed and minimal promoter region of hUTP14a was determined. Differences of the lnciferase activities between different groups were assessed by statistical analysis. Results： The hUTP14a gene promoter reporter construct was correctly cloned and was demonstrated to possess promoter activity. The transcription of hUTP14a was not regulated by P53. The minimal promoter region of h UTP14a gene is located between -203 to -100 of the transcription initiation site. Conclusion： Unlike other P53-interacting proteins such as MDM2, Pirh2 and Cop I which promote P53 degradation and whose transcriptions are regulated by P53, does not hUTP14a transcription form a regulation feedback loop with P 53.

关 键 词：hUTP 14a luciferase activity PROMOTER P5 3 transcription factor 

分 类 号：R730.2[医药卫生—肿瘤]