Hepatitis C Virus Particle Assembly Involves Phosphorylation of NS5A by the c-Abl Tyrosine Kinase

Yamauchi, Shota; Takeuchi, Kenji; Chihara, Kazuyasu; Sun, Xuedong; Honjoh, Chisato; Yoshiki, Hatsumi; Hotta, Hak; Sada, Kiyonao

doi:10.1074/jbc.m115.666859

Cited by 19 publications

(29 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HEK293T cells were cultured in DMEM supplemented with 10% FBS. HCV (J6/JFH1) infection was performed as described previously4445. Infected cells were incubated for 48 h prior to experiments using IFNs.…”

Section: Methodsmentioning

confidence: 99%

“…The absence of targeted gene products was confirmed by immunoblotting. Immunobloting was performed as described previously44. To identify the mutations introduced by the CRISPR/Cas9 system, genomic DNA was extracted using a NucleoSpin Tissue kit (Macherey-Nagel).…”

Section: Methodsmentioning

confidence: 99%

“…Ubiquitin mRNA was used for normalization. To quantify HCV RNA, the standard curve was constructed as described previously44.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

Yamauchi

Takeuchi

Chihara

et al. 2016

Sci Rep

Self Cite

View full text Add to dashboard Cite

Interferon-α (IFN-α) and IFN-λ are structurally distinct cytokines that bind to different receptors, but induce expression of similar sets of genes through Janus kinase (JAK)-signal transducers and activators of transcription (STAT) pathways. The difference between IFN-α and IFN-λ signaling remains poorly understood. Here, using the CRISPR/Cas9 system, we examine the role of STAT1 and STAT2 in the inhibition of hepatitis C virus (HCV) replication by IFN-α and IFN-λ. Treatment with IFN-α increases expression of IFN-stimulated genes (ISGs) such as double-stranded RNA-activated protein kinase (PKR) and decreases viral RNA and protein levels in HCV-infected Huh-7.5 human hepatoma cells. These responses are only partially attenuated by knockout of STAT1 but are abolished by knockout of STAT2. In contrast, the inhibition of HCV replication by IFN-λ is abolished by knockout of STAT1 or STAT2. Microarray analysis reveals that IFN-α but not IFN-λ can induce expression of the majority of ISGs in STAT1 knockout cells. These findings suggest that IFN-α can inhibit HCV replication through a STAT2-dependent but STAT1-independent pathway, whereas IFN-λ induces ISG expression and inhibits HCV replication exclusively through a STAT1- and STAT2-dependent pathway.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

Yamauchi

Takeuchi

Chihara

et al. 2016

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Subsequent reports showed that Abl kinases are also required for infection by several bacteria (e.g. Helicobacter pylori, EPEC, Salmonella enterica, Anaplasma phagocytophilum, Pseudomonas aeruginosa, Chlamydia trachomatis, Mycobacterium tuberculosis) (Bauer et al, 2009;Bruns et al, 2012;Elwell et al, 2008;Jayaswal et al, 2010;Lin et al, 2007;Ly and Casanova, 2009;Muller, 2012;Napier et al, 2011;Pielage et al, 2008;Poppe et al, 2007;Swimm et al, 2004;Tammer et al, 2007) and viruses [vaccinia virus,coxsackievirus,enterovirus 71 (EV71), HIV, hepatitis C virus (HCV) and Ebola virus] (Chen et al, 2007;Coyne and Bergelson, 2006;Garcia et al, 2012;Harmon et al, 2010;Newsome et al, 2006;Reeves et al, 2005;Yamauchi et al, 2015). Microbial pathogens exploit the ability of Abl kinases to regulate cytoskeletal processes in order to achieve efficient internalization, intracellular motility, pedestal formation, cell-to-cell spread, membrane modeling and release (egress).…”

Section: Box 2 Microbial Pathogens Subvert the Function Of Abl Kinasmentioning

confidence: 99%

“…Microbial pathogens exploit the ability of Abl kinases to regulate cytoskeletal processes in order to achieve efficient internalization, intracellular motility, pedestal formation, cell-to-cell spread, membrane modeling and release (egress). Microbial pathogens subvert the function of mammalian Abl kinases by two means: (1) through the deregulation of host cell factors targeted by the Abl kinases, such as actin regulatory proteins (N-WASP, cortactin, Crk, Vav) and GTPases (Rac, Cdc42) and (2) through Abl-mediated phosphorylation of microbial factors required for pathogen entry, motility, and/or release, such as H. pylori CagA (Muller, 2012;Poppe et al, 2007), EPEC Tir (Swimm et al, 2004), vaccinia virus A36R (Newsome et al, 2006;Reeves et al, 2005), Ebola virus VP40 (Garcia et al, 2012) and HCV NS5A (Yamauchi et al, 2015). Mammalian Abl kinases have also been shown to regulate assembly and release of viral particles in the case of HCV (Yamauchi et al, 2015), as well as EV71-induced apoptosis of neuronal cells by targeting cyclin-dependent kinase 5 (CDK5) of the host cell (Chen et al, 2007).…”

Section: Box 2 Microbial Pathogens Subvert the Function Of Abl Kinasmentioning

confidence: 99%

Multifunctional Abl kinases in health and disease

Khatri

Wang

Pendergast

2016

Journal of Cell Science

117

145

View full text Add to dashboard Cite

The Abelson tyrosine kinases were initially identified as drivers of leukemia in mice and humans. The Abl family kinases Abl1 and Abl2 regulate diverse cellular processes during development and normal homeostasis, and their functions are subverted during inflammation, cancer and other pathologies. Abl kinases can be activated by multiple stimuli leading to cytoskeletal reorganization required for cell morphogenesis, motility, adhesion and polarity. Depending on the cellular context, Abl kinases regulate cell survival and proliferation. Emerging data support important roles for Abl kinases in pathologies linked to inflammation. Among these are neurodegenerative diseases and inflammatory pathologies. Unexpectedly, Abl kinases have also been identified as important players in mammalian host cells during microbial pathogenesis. Thus, the use of Abl kinase inhibitors might prove to be effective in the treatment of pathologies beyond leukemia and solid tumors. In this Cell Science at a Glance article and in the accompanying poster, we highlight the emerging roles of Abl kinases in the regulation of cellular processes in normal cells and diverse pathologies ranging from cancer to microbial pathogenesis.

show abstract

The Casein Kinase 2‐Dependent Phosphorylation of NS5A Domain 3 from Hepatitis C Virus Followed by Time‐Resolved NMR Spectroscopy

2016

View full text Add to dashboard Cite

Hepatitis C virus (HCV) chronically affects millions of individuals worldwide. The HCV nonstructural protein 5A (NS5A) plays a critical role in the viral assembly pathway. Domain 3 (D3) of NS5A is an unstructured polypeptide responsible for the interaction with the core particle assembly structure. Casein kinase 2 (CK2) phosphorylates NS5A-D3 at multiple sites that have mostly been predicted and only observed indirectly. In order to identify the CK2-dependent phosphorylation sites, we monitored the reaction between NS5A-D3 and CK2 in vitro by time-resolved NMR. We unambiguously identified four serine residues as substrates of CK2. The apparent rate constant for each site was determined from the reaction curves. Ser408 was quickly phosphorylated, whereas the three other serine residues reacted more slowly. These results provide a starting point from which to elucidate the role of phosphorylation in the mechanisms of viral assembly-and in the modulation of the viral activity-at the molecular level.

show abstract

Hepatitis C Virus Particle Assembly Involves Phosphorylation of NS5A by the c-Abl Tyrosine Kinase

Cited by 19 publications

References 57 publications

STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

Multifunctional Abl kinases in health and disease

The Casein Kinase 2‐Dependent Phosphorylation of NS5A Domain 3 from Hepatitis C Virus Followed by Time‐Resolved NMR Spectroscopy

Contact Info

Product

Resources

About