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

Fan, Yao Shan; Liang, Yi; Liu, Yu

doi:10.1038/s41419-022-04852-3

Cited by 9 publications

(6 citation statements)

References 56 publications

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“…Later on, both ATM and ATR were described as enhancing HBV replication (Kostyusheva et al, 2019), and the ATR/Check1 pathway shown to up-regulate cccDNA formation (Luo et al, 2020a). More recently, DNA-PK was described to increase HBV transcription (Fan et al, 2022). In our analyses, these three kinases were predicted to be activated upon HBV infection, as soon as 2- and up to 7-dpi.…”

Section: Discussionmentioning

confidence: 99%

Deciphering the phospho-signature induced by hepatitis B virus in primary human hepatocytes

Pastor,

Charles,

Belmudes

et al. 2024

Preprint

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Phosphorylation is a major post-translation modification (PTM) of proteins, and small molecules, which is finely tuned by the activity of several hundred kinases and phosphatases. It controls most if not all cellular pathways including anti-viral responses. Accordingly, viruses often induce important changes in the phosphorylation of host factors that can either help or counteract viral replication. Among more than 500 kinases constituting the human kinome only few have been described as important for the Hepatitis B virus (HBV) infectious cycle, and most of them intervene during early or late infectious steps by phosphorylating the viral Core protein (HBc) protein. In addition, scarce information is available on the consequences of HBV infection on the activity of cellular kinases. The objective of this study was to investigate the global impact of HBV infection on the cellular phosphorylation landscape at early after infection. For this, primary human hepatocytes (PHH) were challenged or not with HBV, and a mass spectrometry (MS)-based quantitative phosphoproteomic analysis was conducted two- and seven-days post-infection. The results indicated that while, as expected, HBV infection only minimally modified the cell proteome, important changes were observed on the phosphorylation level of several host proteins at both times points. Gene enrichment and ontology analyses of up- and down- phosphorylated proteins revealed common and distinct signatures induced following infection. In particular, HBV infection resulted in up-phosphorylation of proteins involved in DNA damage signaling and repair, RNA metabolism, in particular splicing, and cytoplasmic cell-signaling. Several down-phosphorylated factors, mostly involved in cell signaling and communication, were also detected. Validation studies performed on some selected up-phosphorylated proteins, revealed that HBV infection induced a DNA damage response characterized by the appearance of 53BP1 foci, whose siRNA-mediated knock-down increased cccDNA levels. In addition, among up-phosphorylated RBPs, SRRM2, a major scaffold of nuclear speckles behaved as antiviral factor. In accordance with these findings, kinase prediction analysis indicated that HBV infection upregulates the activity of major kinases involved in DNA repair. These results strongly suggest that HBV infection triggers an intrinsic anti-viral response composed by DNA repair factors and RBPs that contribute to reduce HBV replication in cell culture models.

show abstract

Section: Discussionmentioning

confidence: 99%

Deciphering the phospho-signature induced by hepatitis B virus in primary human hepatocytes

Pastor,

Charles,

Belmudes

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…Later on, both ATM and ATR were described as enhancing HBV replication ( Kostyusheva et al, 2019 ), and the ATR/Check1 pathway shown to upregulate cccDNA formation ( Luo et al, 2020a ). More recently, DNA-PK was described to increase HBV transcription ( Fan et al, 2022 ). In our analyses, these three kinases were predicted to be activated upon HBV infection, as soon as 2- and up to 7-dpi.…”

Section: Discussionmentioning

confidence: 99%

Deciphering the phospho-signature induced by hepatitis B virus in primary human hepatocytes

Pastor,

Charles,

Belmudes

et al. 2024

Front. Microbiol.

View full text Add to dashboard Cite

Phosphorylation is a major post-translation modification (PTM) of proteins which is finely tuned by the activity of several hundred kinases and phosphatases. It controls most if not all cellular pathways including anti-viral responses. Accordingly, viruses often induce important changes in the phosphorylation of host factors that can either promote or counteract viral replication. Among more than 500 kinases constituting the human kinome only few have been described as important for the hepatitis B virus (HBV) infectious cycle, and most of them intervene during early or late infectious steps by phosphorylating the viral Core (HBc) protein. In addition, little is known on the consequences of HBV infection on the activity of cellular kinases. The objective of this study was to investigate the global impact of HBV infection on the cellular phosphorylation landscape early after infection. For this, primary human hepatocytes (PHHs) were challenged or not with HBV, and a mass spectrometry (MS)-based quantitative phosphoproteomic analysis was conducted 2- and 7-days post-infection. The results indicated that while, as expected, HBV infection only minimally modified the cell proteome, significant changes were observed in the phosphorylation state of several host proteins at both time points. Gene enrichment and ontology analyses of up- and down-phosphorylated proteins revealed common and distinct signatures induced by infection. In particular, HBV infection resulted in up-phosphorylation of proteins involved in DNA damage signaling and repair, RNA metabolism, in particular splicing, and cytoplasmic cell-signaling. Down-phosphorylated proteins were mostly involved in cell signaling and communication. Validation studies carried out on selected up-phosphorylated proteins, revealed that HBV infection induced a DNA damage response characterized by the appearance of 53BP1 foci, the inactivation of which by siRNA increased cccDNA levels. In addition, among up-phosphorylated RNA binding proteins (RBPs), SRRM2, a major scaffold of nuclear speckles behaved as an antiviral factor. In accordance with these findings, kinase prediction analysis indicated that HBV infection upregulates the activity of major kinases involved in DNA repair. These results strongly suggest that HBV infection triggers an intrinsic anti-viral response involving DNA repair factors and RBPs that contribute to reduce HBV replication in cell culture models.

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“…Under the supercoiling stress of transcription (Ma and Wang, 2016), centromeres may produce different types of DNA structures (Kasinathan and Henikoff, 2018) as well as mismatches between repeats. In the centromere associated proteins, both Mre11 and Prkdc can take functions on specific DNA structures (D'Amours and Jackson, 2002;Fan et al, 2022) and Mlh1 is a key player for mismatch repair. The loading of these DNA Frontiers in Genetics frontiersin.org repair proteins to centromeres might be beneficial for the maintenance of centromere stability.…”

Section: Discussionmentioning

confidence: 99%

Dynamic of centromere associated RNAs and the centromere loading of DNA repair proteins in growing oocytes

Yang

Xia

et al. 2023

Front. Genet.

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Mammalian centromeres are generally composed of dispersed repeats and the satellites such as α-satellites in human and major/minor satellites in mouse. Transcription of centromeres by RNA polymerase II is evolutionary conserved and critical for kinetochore assembly. In addition, it has been found that the transcribed satellite RNAs can bind DNA repair proteins such as MRE11 and PRKDC, and excessively expressed satellite RNAs could induce genome instability and facilitate tumorigenesis. During the maturation of female oocyte, centromeres are critical for accurate segregation of homologous chromosomes and sister chromatids. However, the dynamics of oocyte centromere transcription and whether it associated with DNA repair proteins are unknown. In this study, we found the transcription of centromeres is active in growing oocytes but it is silenced when oocytes are fully grown. DNA repair proteins like Mlh1, Mre11 and Prkdc are found associated with the minor satellites and this association can be interfered by RNA polymerase II inhibitor α-amanitin. When the growing oocyte is in vitro matured, Mlh1/Mre11/Prkdc foci would release from centromeres to the ooplasm. If the oocytes are treated with Mre11 inhibitor Mirin, the meiosis resumption of growing oocytes with Mre11 foci can be suppressed. These data revealed the dynamic of centromeric transcription in oocytes and its potential association with DNA repair proteins, which provide clues about how oocytes maintain centromere stability and assemble kinetochores.

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

Cited by 9 publications

References 56 publications

Deciphering the phospho-signature induced by hepatitis B virus in primary human hepatocytes

Deciphering the phospho-signature induced by hepatitis B virus in primary human hepatocytes

Deciphering the phospho-signature induced by hepatitis B virus in primary human hepatocytes

Dynamic of centromere associated RNAs and the centromere loading of DNA repair proteins in growing oocytes

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