Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region

吉颖; 刘竹; 艾青; 朱江; 谢濛宇; 翁华莉; 刘革力; 宋方洲; 卜友泉

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Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region

更新时间：2026-01-09

- Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region
- Journal of Army Medical University Vol. 34, Issue 5, Pages: 391-395(2012)
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  CLC： R394-33;R394.3
- Published：2012
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吉颖,刘竹,艾青,朱江,谢濛宇,翁华莉,刘革力,宋方洲,卜友泉.人HAS3基因核心启动子区域的初步鉴定与分析[J].陆军军医大学学报(原第三军医大学学报),2012,34(05):391-395.

　Ji Ying,Liu Zhu,Ai Qing,Zhu Jiang,Xie Mengyu,Weng Huali,Liu Geli,Song Fangzhou,Bu Youquan.Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(05):391-395.
吉颖,刘竹,艾青,朱江,谢濛宇,翁华莉,刘革力,宋方洲,卜友泉.人HAS3基因核心启动子区域的初步鉴定与分析[J].陆军军医大学学报(原第三军医大学学报),2012,34(05):391-395. DOI：

　Ji Ying,Liu Zhu,Ai Qing,Zhu Jiang,Xie Mengyu,Weng Huali,Liu Geli,Song Fangzhou,Bu Youquan.Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(05):391-395. DOI：

摘要

目的对人HAS3基因的核心启动子区域进行初步鉴定和分析，为深入研究HAS3基因的转录调控奠定基础。方法以前期构建的人HAS3基因启动子荧光素酶报告基因重组体P(-761/-305)为模板，采用PCR介导的基因定点突变技术构建4个不同的系列删除体，并对HAS3启动子区域中的Sp1结合位点和核心启动子元件进行定点突变，构建相应的定点突变重组体；采用荧光素酶双报告基因分析技术检测各重组体的启动子活性。结果构建了P(-761/-569)、P(-563/-305)、P(-490/-305)和P(-433/-305)4个系列删除体和5个定点突变体；启动子活性分析结果表明，P(-761/-569) 无启动子活性，P(-563/-305)、P(-490/-305)和P(-433/-305)均具有较强的启动子活性，Sp1结合位点和核心启动子元件的定点突变可导致HAS3启动子活性的降低。结论 HAS3基因的核心启动子区域主要位于其转录起始位点上游附近-433～-305 bp内，Sp1可能在HAS3的转录调控中起重要作用。

Abstract

Objective To preliminarily identify and characterize the core promoter region for human hyaluronan synthase 3 (HAS3) gene. Methods The previous constructed HAS3 promoter reporter

P(-761/-305)

was used as template to make four different deletion mutants and site-directed mutants by using PCR based site-directed mutagenesis. Dual luciferase reporter assay was used to determine promoter reporter activity. Results Four deletion mutants were constructed

including P(-761/-569)

P(-563/-305)

P(-490/-305) and P(-433/-305). Among the four deletion mutants

P(-761/-569) had no promoter activity whereas P(-563/-305)

P(-490/-305) and P(-433/-305) showed significant promoter activity. Site-directed mutagenesis of Sp1-binding site and core promoter element resulted in a decrease in HAS3 promoter activity. Conclusion The core promoter region of HAS3 gene is mainly located in a 129-bp region (-433 bp to -305 bp) nearby the major transcriptional start site. Sp1 might play an important role in the transcriptional regulation of HAS3 gene.

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