Dual modes of rabies P-protein association with microtubules: a novel strategy to suppress the antiviral response

Moseley, Gregory W.; Lahaye, Xavier; Roth, Daniela; Oksayan, Sibil; Filmer, Richard P.; Rowe, Caitlin; Blondel, Danielle; Jans, David A.

doi:10.1242/jcs.045542

Cited by 69 publications

(118 citation statements)

References 48 publications

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“…In fact, by using a recombinant protein expression system, it was previously demonstrated that P3 inhibits nuclear translocation and DNA binding of STATs (11,17). Further, Marschalek et al (18) showed by using an RABV strain genetically modified to overexpress P2 that this isoform plays an important role in the IFN resistance of the virus.…”

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confidence: 99%

Roles of the Rabies Virus Phosphoprotein Isoforms in Pathogenesis

Okada

Ito

Yamaoka

et al. 2016

J Virol

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Rabies virus (RABV) P gene mRNA encodes five in-frame start codons, resulting in expression of full-length P protein (P1) and N-terminally truncated P proteins (tPs), designated P2, P3, P4, and P5. Despite the fact that some tPs are known as interferon (IFN) antagonists, the importance of tPs in the pathogenesis of RABV is still unclear. In this study, to examine whether tPs contribute to pathogenesis, we exploited a reverse genetics approach to generate CE(NiP)⌬P2-5, a mutant of pathogenic CE(NiP) in which the P gene was mutated by replacing all of the start codons (AUG) for tPs with AUA. We confirmed that while CE(NiP) expresses detectable levels of P2 and P3, CE(NiP)⌬P2-5 has an impaired ability to express these tPs. After intramuscular inoculation, CE(NiP)⌬P2-5 caused significantly lower morbidity and mortality rates in mice than did CE ( Rabies virus (RABV), a member of the genus Lyssavirus of the family Rhabdoviridae, is a zoonotic agent that causes a lethal neurological disease in various mammal species, including humans. After transmission via a bite wound caused by an infected animal, RABV infects peripheral nerves and then spreads to and in the central nervous system (CNS), resulting in severe neurological symptoms with a high case fatality rate of almost 100% (reviewed in reference 1). Due to the absence of an effective cure and insufficient provision of postexposure prophylaxis, approximately 59,000 people die from rabies every year, mainly in developing countries (2). To establish an effective cure and also to develop a novel prophylaxis approach for rabies, it is necessary to understand the molecular mechanisms of the pathogenesis, including immune evasion, of RABV.The phosphoprotein (P protein) of RABV is a multifunctional protein that is indispensable not only for viral replication but also for evasion of host innate immunity. Specifically, this protein plays an essential role in viral RNA synthesis as a cofactor of viral RNA-dependent RNA polymerase (L protein) by bridging nucleoprotein (N protein), which directly binds to viral genomic RNA, and L protein in the ribonucleoprotein complex (reviewed in reference 3). In addition, P protein functions to antagonize the type I interferon (IFN)-mediated antiviral responses by inhibiting both signaling pathways for IFN induction and response (4-12). P protein suppresses activation of interferon regulatory factor 3 (IRF-3), which is an important transcription factor for IFN induction (5, 8). Also, P protein binds to the transcriptional factors signal transducers and activator of transcription 1 (STAT1) and STAT2, which play a key role in the IFN response by activating expression of IFN-stimulated genes (ISGs), and inhibits their nuclear translocation and DNA binding (6,10,11).In RABV-infected cells, mRNA of the P gene is translated from five in-frame start codons by a ribosomal leaky scanning mechanism, resulting in expression of full-length P protein (P1; 297 amino acids) and also less abundant expression of N-terminally truncated P proteins (t...

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confidence: 99%

Roles of the Rabies Virus Phosphoprotein Isoforms in Pathogenesis

Okada

Ito

Yamaoka

et al. 2016

J Virol

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“…Importantly, the results explain for the first time how, through dynamic interactions with MTs, MTAS-dependent MT interaction can facilitate nuclear import rather than resulting in cytoplasmic sequestration and consequently inhibition of nuclear import (47)(48)(49).…”

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confidence: 82%

Mechanism of Microtubule-facilitated “Fast Track” Nuclear Import

Roth

Moseley

Pouton

et al. 2011

Journal of Biological Chemistry

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Although the microtubule (MT) cytoskeleton has been shown to facilitate nuclear import of specific cancer-regulatory proteins including p53, retinoblastoma protein, and parathyroid hormone-related protein (PTHrP), the MT association sequences (MTASs) responsible and the nature of the interplay between MT-dependent and conventional importin (IMP)-dependent nuclear translocation are unknown. Here we used sitedirected mutagenesis, live cell imaging, and direct IMP and MT binding assays to map the MTAS of PTHrP for the first time, finding that it is within a short modular region (residues 82-108) that overlaps with the IMP␤1-recognized nuclear localization signal (residues 66 -108) of PTHrP. Importantly, fluorescence recovery after photobleaching experiments indicated that disruption of the MT network or mutation of the MTAS of PTHrP decreases the rate of nuclear import by 2-fold. Moreover, MTAS functions depend on mutual exclusivity of binding of PTHrP to MTs and IMP␤1 such that, following MTdependent trafficking toward the nucleus, perinuclear PTHrP can be displaced from MTs by IMP␤1 prior to import into the nucleus. This is the first molecular definition of an MTAS that facilitates protein nuclear import as well as the first delineation of the mechanism whereby cargo is transferred directly from the cytoskeleton to the cellular nuclear import apparatus. The results have broad significance with respect to fundamental processes regulating cell physiology/transformation.Nuclear localization of many cancer-regulatory proteins is integral to their function (1-6) in the same way that efficient subcellular trafficking of viruses/viral particles is crucial for pathogenicity (7,8). Nuclear protein import is generally accepted to be independent of the cytoskeleton (9), conventionally being mediated by soluble transport factors (importins (IMPs)), 3 which promote the passage of cargoes bearing nuclear localization signals (NLSs) through the nuclear envelope-localized nuclear pore complex. It is becoming increasingly apparent, however, that specific cancer-related proteins, such as the tumor suppressors p53 (4) and retinoblastoma protein (Rb) (9), and the paracrine/autocrine signaling molecule parathyroid hormone-related protein (PTHrP) (10), rely on the MT network for efficient nuclear import as do certain transcription factors (e.g. STATs 1 and 5) (11,12). PTHrP is a crucial factor in the early stages of development of various tissues as well as in the biology of adult tissues (13,14). Importantly, it is present as a circulating factor in numerous types of tumors (15) with its malignant actions linked to its nuclear localization (2, 3), which significantly is known to be dependent on the IMP family member IMP␤1, as well as MT integrity (10). Specific nuclear localizing proteins, such as PTHrP, appear to possess the ability to interact with MTs for facilitated transport through the cytoplasm prior to conventional IMP-dependent import through the nuclear pore complex into the nucleus (9, 16, 17). However, neither the sequenc...

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“…This unique mechanism of rabies viral subversion of IFN signaling is critical to its pathogenicity, and also suggests novel targets for the development of antiviral drugs or attenuated viruses for vaccine applications [74].…”

Section: Viral Spread To the Cnsmentioning

confidence: 98%

Perspectives in Diagnosis and Treatment of Rabies Viral Encephalitis: Insights from Pathogenesis

et al. 2016

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Rabies viral encephalitis, though one of the oldest recognized infectious disease of humans, remains an incurable, fatal encephalomyelitis, despite advances in understanding of its pathobiology. Advances in science have led us on the trail of the virus in the host, but the sanctuaries in which the virus remains hidden for its survival are unknown. Insights into host-pathogen interactions have facilitated evolving immunologic therapeutic strategies, though we are far from a cure. Most of the present-day knowledge has evolved from in vitro studies using fixed (attenuated) laboratory strains that may not be applicable in the clinical setting. Much remains to be unraveled about this elusive virus. This review attempts to re-examine the current advances in understanding of the pathobiology of the rabies virus that modulate the diagnosis, treatment, and prevention of this fatal disease.

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Dual modes of rabies P-protein association with microtubules: a novel strategy to suppress the antiviral response

Cited by 69 publications

References 48 publications

Roles of the Rabies Virus Phosphoprotein Isoforms in Pathogenesis

Roles of the Rabies Virus Phosphoprotein Isoforms in Pathogenesis

Mechanism of Microtubule-facilitated “Fast Track” Nuclear Import

Perspectives in Diagnosis and Treatment of Rabies Viral Encephalitis: Insights from Pathogenesis

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