title: Cloning and Transcriptional Activity of the Mouse Omi/HtrA2 Gene Promoter.
abstract: HtrA serine peptidase 2 (HtrA2), also named Omi, is a pro-apoptotic protein that exhibits dramatic changes in expression levels in a variety of disorders, including ischemia/reperfusion injury, cancer, and neurodegeneration. In our study, Omi/HtrA2 protein levels were high in the heart, brain, kidney and liver, with elevated heart/brain expression in aging mice. A similar expression pattern was observed at the mRNA level, which suggests that the regulation of Omi/HtrA2 is predominately transcriptional. Promoter binding by transcription factors is the main influencing factor of transcription, and to identify specific promoter elements that contribute to the differential expression of mouse Omi/HtrA2, we constructed truncated Omi/HtrA2 promoter/luciferase reporter vectors and analyzed their relative luciferase activity; it was greatest in the promoter regions at -1205~-838 bp and -146~+93 bp, with the -838~-649 bp region exhibiting negative regulatory activity. Bioinformatics analysis suggested that the Omi/HtrA2 gene promoter contains a CpG island at -709~+37 bp, and eight heat shock transcription factor 1 (HSF1) sites, two Sp1 transcription factor (SP1)sites, one activator protein (AP) site, seven p53 sites, and four YY1 transcription factor(YY1) sites were predicted in the core areas. Furthermore, we found that p53 and HSF1 specifically binds to the Omi/HtrA2 promoter using chromatin immunoprecipitation analysis. These results provide a foundation for understanding Omi/HtrA2 regulatory mechanisms, which could further understanding of HtrA-associated diseases.
authors.affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China. liudanzyh@126.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China. liuxintea@126.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China. wuyecat530@126.com.
authors.forename: Ye
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China. wangwen@ccmu.edu.cn.
authors.forename: Wen
authors.initials: W
authors.lastname: Wang
authors.affiliation: Beijing Key Laboratory of Metabolic Disturbance Related Cardiovascular Disease, Beijing 100069, China. xin.ma@jefferson.edu.|Department of Emergency Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA. xin.ma@jefferson.edu.
authors.forename: Xinliang
authors.initials: X
authors.lastname: Ma
authors.affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China. liuhr2000@126.com.|Beijing Key Laboratory of Metabolic Disturbance Related Cardiovascular Disease, Beijing 100069, China. liuhr2000@126.com.
authors.forename: Huirong
authors.initials: H
authors.lastname: Liu
mesh_terms: D016475:3T3 Cells; D000818:Animals; D018899:CpG Islands; D057809:HEK293 Cells; D000074923:High-Temperature Requirement A Serine Peptidase 2; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D024101:Mitochondrial Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012697:Serine Endopeptidases / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D024101:Mitochondrial Proteins; D014157:Transcription Factors; D012697:Serine Endopeptidases; C000615914:HTRA2 protein, human; D000074923:High-Temperature Requirement A Serine Peptidase 2; C000615915:Htra2 protein, mouse
keywords: Omi/HtrA2; luciferase; promoter; transcriptional activity