Screening and identification of key sites for anti-hepatitis B virus at the carboxyl terminal of APOBEC3B

Chen Yanmeng; Hu Jie; Huang Yao; Yuan Lin; Zhou Xing; Hu Yuan

doi:10.13406/j.cnki.cyxb.001335

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Screening and identification of key sites for anti-hepatitis B virus at the carboxyl terminal of APOBEC3B

更新时间：2024-01-17

- Screening and identification of key sites for anti-hepatitis B virus at the carboxyl terminal of APOBEC3B
- Journal of Chongqing Medical University Vol. 42, Issue 7, Pages: 776-780(2017)
- 作者机构：
  
  1. 重庆医科大学感染性疾病分子生物学教育部重点实验室
  2. 重庆医科大学基础医学院病原微生物教研室
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001335
  CLC： R373
- Published：2017，
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Chen Yanmeng, Hu Jie, Huang Yao, et al. Screening and identification of key sites for anti-hepatitis B virus at the carboxyl terminal of APOBEC3B. [J]. Journal of Chongqing Medical University 42(7):776-780(2017)
DOI：

Chen Yanmeng, Hu Jie, Huang Yao, et al. Screening and identification of key sites for anti-hepatitis B virus at the carboxyl terminal of APOBEC3B. [J]. Journal of Chongqing Medical University 42(7):776-780(2017) DOI： 10.13406/j.cnki.cyxb.001335.

摘要

目的:筛选参与APOBEC3B羧基端脱氨酶及抗乙肝病毒(hepatitis B virus

HBV)活性的重要区域和关键位点。方法:定点突变构建羧基端活性中心附近的3个螺旋区段(α2、α3、α4)的缺失突变体和α4螺旋区内(D316、K320、E321、R327、D328)位点的突变体;Southern blot筛选APOBEC3B抑制HBV复制的重要区段和关键位点;不同DNA变性PCR(differential DNA denaturation PCR

3D-PCR)和克隆测序进行验证。结果:APOBEC3B羧基端脱氨酶活性中心周围的α3和α4螺旋区缺失后

其抗病毒活性消失;D316位点突变后APOBEC3B的脱氨酶活性和抗病毒活性消失。结论:APOBEC3B羧基端的D316位点是APOBEC3B脱氨酶和抗病毒的关键位点。

Abstract

Objective:To screen the important domains and key sites involved in deaminase and anti-hepatitis B virus（HBV）activity at the carboxyl terminal of APOBEC3 B. Methods:We mapped three alpha helices regions（α2

α3

α4）nearby the active site and sites at alpha 4（D316

K320

E321

R327

D328）by site-directed mutagenesis to test whether the mutations alter anti-HBV function of APOBEC3 B. Results:The α3 and α4 regions around the active site of deaminase at the carboxyl terminal of APOBEC3 B were important for anti-HBV;D316 site at the carboxyl terminal of APOBEC3 B was critical for inhibition of HBV depending on its deaminase activity. Conclusion:D316 is catalytically active and capable of restricting.

关键词

APOBEC3B脱氨基3D-PCRD316

Keywords

APOBEC3BDeamination3D-PCRD316

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