PRKDC promotes hepatitis B virus transcription through enhancing the binding of RNA Pol II to cccDNA

作者全名："Fan, Yao; Liang, Yi; Liu, Yu; Fan, Hui"

作者地址："[Fan, Yao; Liang, Yi; Liu, Yu; Fan, Hui] Chongqing Med Univ, Chinese Minist Educ, Key Lab Mol Biol Infect Dis Designated, Chongqing 400016, Peoples R China; [Fan, Yao] Southwest Med Univ, Affiliated Tradit Chinese Med Hosp, Luzhou 646000, Peoples R China"

通信作者："Fan, H (通讯作者)，Chongqing Med Univ, Chinese Minist Educ, Key Lab Mol Biol Infect Dis Designated, Chongqing 400016, Peoples R China."

来源：CELL DEATH & DISEASE

ESI学科分类：MOLECULAR BIOLOGY & GENETICS

WOS号：WOS:000787327700004

JCR分区：Q1

影响因子：9

年份：2022

卷号：13

期号：4

开始页：　

结束页：　

文献类型：Article

关键词：　

摘要："Hepatitis B virus infection remains a major health problem worldwide due to its high risk of liver failure and hepatocellular carcinoma. Covalently closed circular DNA (cccDNA), which is present as an individual minichromosome, serves as the template for transcription of all viral RNAs and pla ays critical role in viral persistence. Therefore, there is an urgent need to gain broader insight into the transcription regulation of cccDNA. Here, we combined a modified Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) with an engineered ascorbate peroxidase 2 (APEX2) to identify cccDNA associated proteins systematically in living cells. By functional screening, we verified that protein kinase, DNA-activated, catalytic subunit (PRKDC) was an effective activator of HBV cccDNA transcription in HBV-infected HepG2-NTCP cells and primary human hepatocytes. Mechanismly, PRKDC interacted with POLR2A and POLR2B, the two largest subunits of RNA polymerase II (Pol II) and recruited Pol II to HBV cccDNA minichromosome in a kinase-dependent manner. PRKDC knockdown or inhibitor treatment significantly decreased the enrichment of POLR2A and POLR2B on cccDNA, as well as reducing the levels of cccDNA associated Pol II Ser5 and Ser2 phosphorylation, which eventually inhibited the HBV cccDNA activity. Collectively, these findings give us new insights into cccDNA transcription regulation, thus providing new potential targets for HBV treatment in patients."

基金机构：high-level young science and technology talent cultivation plan (Chongqing Medical University)

基金资助正文：This work was supported by the financial assistance provided by the high-level young science and technology talent cultivation plan (Chongqing Medical University).