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The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication

We recently reported that the PPIase Par14 and Par17 encoded by PIN4 upregulate HBV replication in an HBx-dependent manner by binding to conserved arginine–proline (RP) motifs of HBx. HBV core protein (HBc) has a conserved (133)RP(134) motif; therefore, we investigated whether Par14/Par17 bind to HB...

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Autores principales: Saeed, Umar, Piracha, Zahra Zahid, Kwon, Hyeonjoong, Kim, Jumi, Kalsoom, Fadia, Chwae, Yong-Joon, Park, Sun, Shin, Ho-Joon, Lee, Hyun Woong, Lim, Jin Hong, Kim, Kyongmin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8713550/
https://www.ncbi.nlm.nih.gov/pubmed/34970249
http://dx.doi.org/10.3389/fmicb.2021.795047
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author Saeed, Umar
Piracha, Zahra Zahid
Kwon, Hyeonjoong
Kim, Jumi
Kalsoom, Fadia
Chwae, Yong-Joon
Park, Sun
Shin, Ho-Joon
Lee, Hyun Woong
Lim, Jin Hong
Kim, Kyongmin
author_facet Saeed, Umar
Piracha, Zahra Zahid
Kwon, Hyeonjoong
Kim, Jumi
Kalsoom, Fadia
Chwae, Yong-Joon
Park, Sun
Shin, Ho-Joon
Lee, Hyun Woong
Lim, Jin Hong
Kim, Kyongmin
author_sort Saeed, Umar
collection PubMed
description We recently reported that the PPIase Par14 and Par17 encoded by PIN4 upregulate HBV replication in an HBx-dependent manner by binding to conserved arginine–proline (RP) motifs of HBx. HBV core protein (HBc) has a conserved (133)RP(134) motif; therefore, we investigated whether Par14/Par17 bind to HBc and/or core particles. Native agarose gel electrophoresis (NAGE) and immunoblotting and co-immunoprecipitation were used. Chromatin immunoprecipitation from HBV-infected HepG2-hNTCP-C9 cells was performed. NAGE and immunoblotting revealed that Par14/Par17 bound to core particles and co-immunoprecipitation revealed that Par14/Par17 interacted with core particle assembly-defective, and dimer-positive HBc-Y132A. Thus, core particles and HBc interact with Par14/Par17. Par14/Par17 interacted with the HBc (133)RP(134) motif possibly via substrate-binding E46/D74 and E71/D99 motifs. Although Par14/Par17 dissociated from core particles upon heat treatment, they were detected in 0.2 N NaOH-treated opened-up core particles, demonstrating that Par14/Par17 bind outside and inside core particles. Furthermore, these interactions enhanced the stabilities of HBc and core particles. Like HBc-Y132A, HBc-R133D and HBc-R133E were core particle assembly-defective and dimer-positive, demonstrating that a negatively charged residue at position 133 cannot be tolerated for particle assembly. Although positively charged R133 is solely important for Par14/17 interactions, the (133)RP(134) motif is important for efficient HBV replication. Chromatin immunoprecipitation from HBV-infected cells revealed that the S19 and E46/D74 residues of Par14 and S44 and E71/D99 residues of Par17 were involved in recruitment of (133)RP(134) motif-containing HBc into cccDNA. Our results demonstrate that interactions of HBc, Par14/Par17, and cccDNA in the nucleus and core particle–Par14/Par17 interactions in the cytoplasm are important for HBV replication.
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spelling pubmed-87135502021-12-29 The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication Saeed, Umar Piracha, Zahra Zahid Kwon, Hyeonjoong Kim, Jumi Kalsoom, Fadia Chwae, Yong-Joon Park, Sun Shin, Ho-Joon Lee, Hyun Woong Lim, Jin Hong Kim, Kyongmin Front Microbiol Microbiology We recently reported that the PPIase Par14 and Par17 encoded by PIN4 upregulate HBV replication in an HBx-dependent manner by binding to conserved arginine–proline (RP) motifs of HBx. HBV core protein (HBc) has a conserved (133)RP(134) motif; therefore, we investigated whether Par14/Par17 bind to HBc and/or core particles. Native agarose gel electrophoresis (NAGE) and immunoblotting and co-immunoprecipitation were used. Chromatin immunoprecipitation from HBV-infected HepG2-hNTCP-C9 cells was performed. NAGE and immunoblotting revealed that Par14/Par17 bound to core particles and co-immunoprecipitation revealed that Par14/Par17 interacted with core particle assembly-defective, and dimer-positive HBc-Y132A. Thus, core particles and HBc interact with Par14/Par17. Par14/Par17 interacted with the HBc (133)RP(134) motif possibly via substrate-binding E46/D74 and E71/D99 motifs. Although Par14/Par17 dissociated from core particles upon heat treatment, they were detected in 0.2 N NaOH-treated opened-up core particles, demonstrating that Par14/Par17 bind outside and inside core particles. Furthermore, these interactions enhanced the stabilities of HBc and core particles. Like HBc-Y132A, HBc-R133D and HBc-R133E were core particle assembly-defective and dimer-positive, demonstrating that a negatively charged residue at position 133 cannot be tolerated for particle assembly. Although positively charged R133 is solely important for Par14/17 interactions, the (133)RP(134) motif is important for efficient HBV replication. Chromatin immunoprecipitation from HBV-infected cells revealed that the S19 and E46/D74 residues of Par14 and S44 and E71/D99 residues of Par17 were involved in recruitment of (133)RP(134) motif-containing HBc into cccDNA. Our results demonstrate that interactions of HBc, Par14/Par17, and cccDNA in the nucleus and core particle–Par14/Par17 interactions in the cytoplasm are important for HBV replication. Frontiers Media S.A. 2021-12-14 /pmc/articles/PMC8713550/ /pubmed/34970249 http://dx.doi.org/10.3389/fmicb.2021.795047 Text en Copyright © 2021 Saeed, Piracha, Kwon, Kim, Kalsoom, Chwae, Park, Shin, Lee, Lim and Kim. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Saeed, Umar
Piracha, Zahra Zahid
Kwon, Hyeonjoong
Kim, Jumi
Kalsoom, Fadia
Chwae, Yong-Joon
Park, Sun
Shin, Ho-Joon
Lee, Hyun Woong
Lim, Jin Hong
Kim, Kyongmin
The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication
title The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication
title_full The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication
title_fullStr The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication
title_full_unstemmed The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication
title_short The HBV Core Protein and Core Particle Both Bind to the PPiase Par14 and Par17 to Enhance Their Stabilities and HBV Replication
title_sort hbv core protein and core particle both bind to the ppiase par14 and par17 to enhance their stabilities and hbv replication
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8713550/
https://www.ncbi.nlm.nih.gov/pubmed/34970249
http://dx.doi.org/10.3389/fmicb.2021.795047
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