干扰素刺激应答元件介导IFN-α-2b对IFP35的转录调控  

An Interferon Stimulate Response Element(ISRE) Mediates IFN-α-2b Induced IFP35 Upregulation

作　　者：杨炜[1] 谈娟[1] 乔文涛[1] 耿运琪[1] 

机构地区：[1]南开大学分子微生物与技术教育部重点实验室,天津300071

出　　处：《南开大学学报（自然科学版）》2012年第5期72-75,81,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家自然科学基金(30900068,31070135)

摘　　要：IFP35为分子量35 kDa的干扰素诱导蛋白,可以被Ⅰ型干扰素诱导表达,但是其受干扰素调控机制未知.为了阐明Ⅰ型干扰素对IFP35的转录调控机制,克隆了IFP35基因翻译起始点上游242 bp(-359～-118)的序列,将之置于萤火虫荧光素酶基因上游,构建报告质粒pGL-359,瞬时转染实验证实,该段序列具有启动子活性;序列分析表明其为GC box型启动子,-181～-170 bp序列符合干扰素刺激应答元件(interferon stimulate response element,ISRE)特征.用IFN-α-2b处理细胞后,pGL-359活性可提高达3.1倍,但突变pGL-359上的ISRE特征序列后,该突变启动子不再响应IFN-α-2b刺激.随后的凝胶电泳阻滞实验发现,用IFN-α-2b处理HeLa细胞后,其核蛋白可与IFP35启动子上的ISRE元件形成明显的阻滞带,表明ISRE介导了IFN-α-2b对IFP35的转录调控.IFP35 is a type I interferon induced protein with a mass of 35 kDa. The promoter of IFP35 （pGL-359） was cloned to elucidate how IFP35 is regulated by type I interferon. HeLa cells were transfected with pGL-359 and were stimulated with IFN-a-2b. The activity of pGL-359 was obviously activated upon harboring the mutated ISRE Completely lost IFN-a-2b stimulation. However, pGL-359mISRE luciferase activity in response to IFN-a-2b. Addi- tionally, electrophoretic mobility-shift assay revealed that the ISRE of IFP35 promoter formed complex with the nuclear extract of HeLa cells that were treated with IFN-a-2b, which demonstrated further that ISRE is responsible for the IFN-a-2b-mediated upregulation of IFP35.

关 键 词：IFP35 干扰素 干扰素刺激应答元件 

分 类 号：Q753[生物学—分子生物学]