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ENPP1 inhibits the transcription activity of the hepatitis B virus pregenomic promoter by upregulating the acetylation of LMNB1

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Abstract

Current therapies for hepatitis B virus (HBV) infection can slow disease progression but cannot cure the infection, as it is difficult to eliminate or permanently silence HBV covalently closed circular DNA (cccDNA). The interaction between host factors and cccDNA is essential for their formation, stability, and transcriptional activity. Here, we focused on the regulatory role of the host factor ENPP1 and its interacting transcription factor LMNB1 in HBV replication and transcription to better understand the network of host factors that regulate HBV, which may facilitate the development of new antiviral drugs. Overexpression of ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) in Huh7 cells decreased HBV pregenomic RNA (pgRNA) and hepatitis B core antigen (HBcAg) expression levels, whereas knockdown of ENPP1 increased them. A series of HBV promoter and mutant plasmids were constructed, and a luciferase reporter assay showed that overexpression of ENPP1 caused inhibition of the HBV promoter and its mutants. A DNA pull-down assay showed that lamin B1 (LMNB1), but not ENPP1, interacts directly with the HBV enhancer II/ basic core promoter (EnhII/BCP). ZDOCK and PyMOL software were used to predict the interaction of ENPP1 with LMNB1. Overexpression of LMNB1 inhibited the activity of the HBV promoter and its mutant. The acetylation levels at the amino acids 111K, 261K, and 483K of LMNB1 were reduced compared to the control, and an LMNB1 acetylation mutant containing 111R, 261Q, 261R, 483Q, and 483R showed increased promoter activity. In summary, ENPP1 together with LMNB1 increased the acetylation level at 111K and 261K, and LMNB1 inhibited the activity of HBV promoter and downregulated the expression of pregenomic RNA and HBcAg. Our follow-up studies will investigate the expression, clinical significance, and relevance of ENPP1 and LMNB1 in HBV patient tissues, explore the effect of LMNB1 on post-transcriptional progression, and examine whether ENPP1 can reduce cccDNA levels in the nucleus.

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Data availability

All data, models, and code generated or used during the study appear in the submitted article.

Abbreviations

cccDNA:

Covalently closed circular DNA

cGAMP:

Cyclic GMP-AMP

ENPP1:

Ectonucleotide pyrophosphatase/phosphodiesterase 1

EnhII / BCP:

Enhancer II/ basic core promoter

HBcAg:

Hepatitis B core antigen

HBV:

Hepatitis B virus

HCV:

Hepatitis C virus

HRP:

Horseradish peroxidase

IFN-a:

Interferon a

LMNB1:

Lamin B1

NAs:

Nucleotide analogues

PEG-IFN:

Pegylated interferon

pgRNA:

Pregenomic RNA

PVDF:

Polyvinylidene fluoride

SDS-PAGE:

SDS polyacrylamide gel electrophoresis

siRNA:

Small interfering RNA

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Funding

This study was supported by the Key Program, co-sponsored by the province and ministry (222301420071), the General Project of the Henan Natural Science Foundation (222300420572, 222102310149), and the Youth Program of the National Natural Science Foundation of China (82104641).

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Author notes

  1. Xinping Ma and Yuan Li have contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, Zhengzhou University People’s Hospital, Henan Provincial People’s Hospital, Zhengzhou, 450003, Henan, China

    Xinping Ma, Yingli Huang, Xiaofang Li, Shuangyin Han, Hui Ding & Suofeng Sun

  2. Department of Traditional Chinese Medicine, The Third Affiliated Hospital Affiliated of Henan University of Traditional Chinese Medicine, Zhengzhou, 450003, Henan, China

    Yuan Li

  3. Department of Gastroenterology, School of Clinical Medicine, Henan Provincial People’s Hospital, Henan University, Zhengzhou, 450003, Henan, China

    Huihui Zhu

  4. Xinxiang Medical University, Xinxiang, 453000, Henan, China

    Kai Lu

  5. The department of infectious diseases, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, 450000, Henan, China

    Xinping Ma

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  1. Xinping Ma
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Correspondence to Hui Ding or Suofeng Sun.

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Ma, X., Li, Y., Zhu, H. et al. ENPP1 inhibits the transcription activity of the hepatitis B virus pregenomic promoter by upregulating the acetylation of LMNB1. Arch Virol 169, 36 (2024). https://doi.org/10.1007/s00705-023-05949-6

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