Heat Shock Protein 70 Inhibits the Activity of Influenza A Virus Ribonucleoprotein and Blocks the Replication of Virus In Vitro and In Vivo

Li, Gang; Zhang, Junjie; Tong, Xiaomei; Liu, Wenjun; Ye, Xin

doi:10.1371/journal.pone.0016546

Cited by 65 publications

(64 citation statements)

References 62 publications

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“…The mouse anti-Hsp70 MAb (SPA-810) and rabbit polyclonal anti-Hsp70 (SPA-812) antibody were obtained from Stressgen, and the anti-␣-tubulin MAb (N356) was from Amersham. Quercetin (Qct; Q4951) and the rabbit polyclonal anti-actin (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33) antibody were obtained from Sigma. The mouse anti-green fluorescent protein (GFP) MAb (11814460001) was purchased from Roche.…”

Section: Methodsmentioning

confidence: 99%

“…An increase of its expression confers to cells a protection against rotavirus (2), vesicular stomatitis virus (9), respiratory syncytial virus (41), and influenza virus (16) infections. In the latter case, it has been reported that Hsp70, which interacts with the ribonucleoprotein complex, interferes with the polymerase activity and negatively regulates viral transcription and replication (16,22). Interestingly, Hsp70 was found to have both positive and negative regulatory effects on the RNA replication of flock house virus, a nodavirus (39), highlighting the complexity of this particular virus-chaperone interaction.…”

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confidence: 99%

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Hsp70 Protein Positively Regulates Rabies Virus Infection

Lahaye

Vidy

Fouquet

et al. 2012

J Virol

123

105

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The Hsp70 chaperone plays a central role in multiple processes within cells, including protein translation, folding, intracellular trafficking, and degradation. This protein is implicated in the replication of numerous viruses. We have shown that rabies virus infection induced the cellular expression of Hsp70, which accumulated in Negri body-like structures, where viral transcription and replication take place. In addition, Hsp70 is present in both nucleocapsids purified from infected cells and in purified virions. Hsp70 has been shown to interact with the nucleoprotein N. The downregulation of Hsp70, using specific chaperone inhibitors, such as quercetin or RNA interference, resulted in a significant decrease of the amount of viral mRNAs, viral proteins, and virus particles. These results indicate that Hsp70 has a proviral function during rabies virus infection and suggest that Hsp70 is involved in at least one stage(s) of the viral life cycle, such as viral transcription, translation, and/or production. The mechanism by which Hsp70 controls viral infection will be discussed.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Hsp70 Protein Positively Regulates Rabies Virus Infection

Lahaye

Vidy

Fouquet

et al. 2012

J Virol

123

105

View full text Add to dashboard Cite

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“…The expression plasmids for the PA, PB1, and PB2 genes from A/WSN/33 virus were generated by cloning into the pcDNA3-FLAG vector as described previously (47). pHH21-cNS-Luc was generated by cloning the cRNA promoter of NS with luciferase into pHH21 as described previously (18). All constructs were verified by DNA sequencing.…”

Section: Cells Viruses and Antibodies Madin-darby Canine Kidney (Mmentioning

confidence: 99%

“…The resulting RNA preparations were treated with RNase-free DNase I (Takara, Tokyo, Japan) according to the manufacturer's protocol. One microgram of total RNA was used for cDNA synthesis with the oligo(dT) 18 primer, followed by real-time PCR quantification. Real time quantitative PCR was performed with SYBR Premix Ex Taq (TaKaRa, Japan) on a Corbett 6200 real-time detection system (Corbett, Australia) to investigate the expression level of luciferase in luciferase assay (54).…”

Section: Nuclear and Cytoplasmic Mrna Quantification By Real-time Revmentioning

confidence: 99%

Identification and Characterization of Three Novel Nuclear Export Signals in the Influenza A Virus Nucleoprotein

Liu

Cao

et al. 2012

J Virol

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The nuclear export of the influenza A virus ribonucleoprotein (vRNP) is crucial for virus replication. As a major component of the vRNP, nucleoprotein (NP) alone can also be shuttled out of the nucleus by interacting with chromosome region maintenance 1 (CRM1) and is therefore hypothesized to promote the nuclear export of the vRNP. In the present study, three novel nuclear export signals (NESs) of the NP-NES1, NES2, and NES3-were identified as being responsible for mediating its nuclear export. The nuclear export of NES3 was CRM1 dependent, whereas that of NES1 or NES2 was CRM1 independent. Inactivation of these NESs led to an overall nuclear accumulation of NP. Mutation of all three NP-NESs significantly impaired viral replication. Based on structures of influenza virus NP oligomers, these three hydrophobic NESs are found present on the surface of oligomeric NPs. Functional studies indicated that oligomerization is also required for nuclear export of NP. Together, these results suggest that the nuclear export of NP is important for virus replication and relies on its NESs and oligomerization.T he influenza A virus genome consists of eight negative-sense single-stranded RNA segments (vRNA) (17). Each vRNA segment is associated with multiple copies of the viral nucleoprotein (NP) and three polymerase subunits (PA, PB1, and PB2), forming the viral ribonucleoprotein (vRNP) complex. During an early stage of infection, the vRNPs are released into the cytoplasm from virions following fusion of the viral membrane and endosomal membrane (41). Subsequently, the incoming vRNPs are transported into the nucleus, where viral genome replication and transcription occur (41).One of the determinants for vRNP nuclear import is NP (8, 28, 44, 49, 52, 53), the major protein in the vRNP structure. Thus far, two nuclear localization signals (NLSs) and a nuclear accumulation signal (NAS) have been identified in NP. The stronger NLS is an unconventional signal located in the N-terminal basic region (between residues 3 and 13) of NP (28, 44), and the weaker signal, a classical bipartite NLS, is located between residues 198 and 216 (49). The NAS-spanning residues (327 to 345) were identified through analyses of mutants that lacked both of the NLSs but still exhibited partial nuclear distribution (9).At a late stage of virus infection, the vRNPs exit the nucleus to assemble and bud from the apical plasma membrane of polarized cells (3). The trafficking of the vRNPs into and out of the nucleus is a tightly regulated process (7). The nuclear export of progeny vRNPs is mediated by the CRM1 cellular export receptor (12,23,29,48). Nuclear export protein (NEP; formerly referred to as the NS2 protein), which possesses a nuclear export signal (NES), and the matrix protein (M1) of influenza A virus are deemed responsible for directing export of the vRNPs (29, 31). This process is regulated by the Raf/MEK/ERK pathway (32), which is stimulated by the membrane association of influenza virus hemagglutinin (HA) (25). Interestingly, exogenously expresse...

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“…Several viruses in fact stimulate a major increase in Hsps toward this end, such as human papillomavirus (Song et al 2010), adenovirus (Glotzer et al 2000;Madara et al 2005), polyomaviruses, and dengue viruses (Reyes-Del Valle et al 2005). Other viruses, however, suffer a reduction in replication when host Hsps are high, such as human immunodeficiency virus (HIV) and influenza A, where Hsp70 inhibits viral gene expression and replication (Kumar et al 2011;Li et al 2011). It is therefore not surprising that some viruses known for inducing chronic fatigue and malaise act to limit the host stress response, including influenza A, West Nile virus, herpes simplex virus, and hepatitis C. Hosts, on the other hand, can respond to viral infections through several defensive maneuvers, including generation of a fever, immunological defense, interferon production, and reduction of protein synthesisincluding reduced Hsp synthesis-which set hosts up for adverse symptoms.…”

Section: Introductionmentioning

confidence: 99%