HIV-1 B、C亚型Tat第41位赖氨酸对HIV-1转录活化的影响  

Differential transactivation of HIV-1 subtypes B and C by Tat lysine 41

作　　者：刘瑜 胡月 孔晓红[2] Liu Yu;Hu Yue;Kong Xiaohong(Clinical Laboratory,Tianjin Medical University Chu Hsien-I Memorial Hospital,Tianjin Institute of Endocrinology,NHC Key Laboratory of Hormones and Development(Tianjin Medical University),Tianjin Key Laboratory of Metabolic Diseases,Tianjin 300134,China;Laboratory of Medical Molecular Virology,School of Medicine,Nankai University,Tianjin 300071,China)

机构地区：[1]天津医科大学朱宪彝纪念医院检验科,天津市内分泌研究所,国家卫生健康委员会激素与发育重点实验室,天津市代谢性疾病重点实验室,300134 [2]南开大学医学院分子病毒学实验室,天津300071

出　　处：《中华实验和临床病毒学杂志》2020年第4期406-410,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：天津医科大学朱宪彝纪念医院科研基金(2018RC04);国家自然科学基金面上项目(81571987)。

摘　　要：目的对反式转录激活因子Tat第41位赖氨酸(K41)在HIV-1转录活化过程中的作用进行探讨,并对B、C亚型HIV-1中Tat K41的作用进行比较。方法通过荧光素酶活性实验检测野生型Tat、突变型Tat K41A、Tat K41R对HIV-1转录活性的影响,并比较B、C亚型HIV-1中Tat K41作用的差异;通过免疫沉淀(immunoprecipitation,IP)实验检测B、C亚型HIV-1中Tat K41是否影响Tat与SIRT1的结合;通过Western blot检测细胞核内B、C亚型HIV-1的Tat K41是否影响p65 K310的乙酰化水平。结果与B亚型HIV-1不同,C亚型HIV-1的Tat K41突变后显著降低荧光素酶活性,减弱C亚型Tat与SIRT1的结合,降低细胞核内p65 K310ac的水平。结论C亚型Tat K41在HIV-1转录中发挥重要作用。Objective To investigate the effect of trans-activation of transcription(Tat)lysine 41 on HIV-1 transcription and compare the effect of HIV-1 subtypes B and C.Methods The effects of wildtype Tat,Tat K41A and Tat K41R mutants on HIV-1 transcription were assessed by luciferase assay,and the differences of Tat K41 effect were compared in subtypes B and C viruses.Next,immunoprecipitation(IP)assay was used to detect whether Tat K41 of HIV-1 clades B and C affects the interaction between Tat and SIRT1.Finally,Western blot was used to detect whether K41 of Tat B and Tat C affects the acetylation level of p65 K310 in the nucleus.Results Mutants of subtype C Tat K41 significantly inhibited Tat transactivation,reduced the interaction between SIRT1 and Tat C and decreased the level of p65 K310ac in the nucleus compared with wildtype of subtype C Tat,but there were no significant changes in subtype B Tat.Conclusions Subtype C Tat K41 plays a vital role in HIV-1 transcription.

关 键 词：人免疫缺陷病毒 反式转录激活因子 转录 B亚型 C亚型 

分 类 号：R373[医药卫生—病原生物学]