The C-Terminal End of Parainfluenza Virus 5 NP Protein Is Important for Virus-Like Particle Production and M-NP Protein Interaction

Schmitt, Phuong Tieu; Ray, Greeshma; Schmitt, Anthony P.

doi:10.1128/jvi.01885-10

Cited by 24 publications

(29 citation statements)

References 54 publications

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“…After an additional 18 h, cell and medium fractions were collected. VLPs from the culture medium fractions were pelleted through 20% sucrose cushions, resuspended, floated to the tops of sucrose flotation gradients, pelleted again, and then resuspended in SDS-PAGE loading buffer containing 2.5% (wt/vol) dithiothreitol, as described previously (34). Cell lysate preparation and immunoprecipitation of proteins from the cell lysate fraction were carried out as described previously (23).…”

Section: Methodsmentioning

confidence: 99%

Matrix Proteins of Nipah and Hendra Viruses Interact with Beta Subunits of AP-3 Complexes

Sun

McCrory

Khaw

et al. 2014

J Virol

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Paramyxoviruses and other negative-strand RNA viruses encode matrix proteins that coordinate the virus assembly process. The matrix proteins link the viral glycoproteins and the viral ribonucleoproteins at virus assembly sites and often recruit host machinery that facilitates the budding process. Using a co-affinity purification strategy, we have identified the beta subunit of the AP-3 adapter protein complex, AP3B1, as a binding partner for the M proteins of the zoonotic paramyxoviruses Nipah virus and Hendra virus. Binding function was localized to the serine-rich and acidic Hinge domain of AP3B1, and a 29-amino-acid Hingederived polypeptide was sufficient for M protein binding in coimmunoprecipitation assays. Virus-like particle (VLP) production assays were used to assess the relationship between AP3B1 binding and M protein function. We found that for both Nipah virus and Hendra virus, M protein expression in the absence of any other viral proteins led to the efficient production of VLPs in transfected cells, and this VLP production was potently inhibited upon overexpression of short M-binding polypeptides derived from the Hinge region of AP3B1. Both human and bat (Pteropus alecto) AP3B1-derived polypeptides were highly effective at inhibiting the production of VLPs. VLP production was also impaired through small interfering RNA (siRNA)-mediated depletion of AP3B1 from cells. These findings suggest that AP-3-directed trafficking processes are important for henipavirus particle production and identify a new host protein-virus protein binding interface that could become a useful target in future efforts to develop small molecule inhibitors to combat paramyxoviral infections. IMPORTANCEHenipaviruses cause deadly infections in humans, with a mortality rate of about 40%. Hendra virus outbreaks in Australia, all involving horses and some involving transmission to humans, have been a continuing problem. Nipah virus caused a large outbreak in Malaysia in 1998, killing 109 people, and smaller outbreaks have since occurred in Bangladesh and India. In this study, we have defined, for the first time, host factors that interact with henipavirus M proteins and contribute to viral particle assembly. We have also defined a new host protein-viral protein binding interface that can potentially be targeted for the inhibition of paramyxovirus infections.

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Section: Methodsmentioning

confidence: 99%

Matrix Proteins of Nipah and Hendra Viruses Interact with Beta Subunits of AP-3 Complexes

Sun

McCrory

Khaw

et al. 2014

J Virol

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“…Interestingly, the degree of neuroattenuation imparted by the N/M gene combination was substantially greater than an additive effect of the two genes alone (25% and 40%, respectively), suggesting a role of an N/M interaction in virus virulence. Given that the paramyxovirus M protein physically interacts with the N protein to facilitate virus assembly via associating the RNP with areas of the host cell membrane containing the viral glycoproteins (2,22,26,37,38,44), it is noteworthy that in comparison to parental r88, the N/M gene combination chimeric virus grew to 100-fold-lower titers in vivo. Whether efficient viral assembly or budding is impaired in r88 viruses expressing the rJL N and/or M genes requires further investigation.…”

Section: Vol 85 2011 Genetic Determinants Of Mumps Virus Neurovirulmentioning

confidence: 99%

“…The HN glycoprotein mediates attachment of the virus to its cellular receptor (sialic acid) and, together with the F glycoprotein, mediates fusion of the virion membrane with the target cell membrane. Based on studies of related paramyxoviruses, the M protein is involved in assembling the RNP at the plasma membrane in regions containing the HN and F glycoproteins, presumably via interacting with the carboxy-terminal tail portion of the RNP-associated N protein and the cytoplasmic tails of the glycoproteins (11,16,38). The SH protein itself is not required for virus replication but is believed to serve luxury functions, such as exhibiting antiapoptotic activity (43, 47).…”

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confidence: 99%

Gene-Specific Contributions to Mumps Virus Neurovirulence and Neuroattenuation

Sauder

Zhang

Ngo

et al. 2011

J Virol

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Mumps virus (MuV) is highly neurotropic and was the leading cause of aseptic meningitis in the WesternHemisphere prior to widespread use of live attenuated MuV vaccines. Due to the absence of markers of virus neuroattenuation and neurovirulence, ensuring mumps vaccine safety has proven problematic, as demonstrated by the occurrence of aseptic meningitis in recipients of certain vaccine strains. Here we examined the genetic basis of MuV neuroattenuation and neurovirulence by generating a series of recombinant viruses consisting of combinations of genes derived from a cDNA clone of the neurovirulent wild-type 88-1961 strain (r88) and from a cDNA clone of the highly attenuated Jeryl Lynn vaccine strain (rJL). Testing of these viruses in rats demonstrated the ability of several individual rJL genes and gene combinations to significantly neuroattenuate r88, with the greatest effect imparted by the rJL nucleoprotein/matrix protein combination. Interestingly, no tested combination of r88 genes, including the nucleoprotein/matrix protein combination, was able to convert rJL into a highly neurovirulent virus, highlighting mechanistic differences between processes involved in neuroattenuation and neurovirulence.Mumps virus (MuV) is a paramyxovirus belonging to the Rubulavirus genus. The nonsegmented negative-sense genome of 15,384 nucleotides contains the following seven transcription units: the nucleoprotein (N), the V/phosphoprotein (V/P/ I), the matrix protein (M), the fusion protein (F), the small hydrophobic protein (SH), the hemagglutinin-neuraminidase protein (HN), and the large protein (L) genes. Each gene encodes a single protein, with the exception of the V/P/I gene (conventionally referred to as the P gene), which gives rise to additional mRNA species as a result of the cotranscriptional insertion of nontemplated G nucleotides between positions 461 and 466, the so-called editing site. Faithful transcription of the gene produces the V protein, whereas insertion of two G residues within the editing site produces an mRNA encoding the P protein, and insertion of four G residues produces an mRNA encoding the I protein, analogous to the W protein identified in other paramyxoviruses (9,20,28). The roles of the viral proteins in the life cycle of paramyxoviruses have been well described in the literature (9,12,20). Briefly, the N protein encapsidates the nascent viral RNA as it is being synthesized, forming a ribonucleoprotein (RNP) complex, which serves as the substrate for the viral RNA-dependent RNA polymerase (RdRp), formed by the P and L proteins. The HN glycoprotein mediates attachment of the virus to its cellular receptor (sialic acid) and, together with the F glycoprotein, mediates fusion of the virion membrane with the target cell membrane. Based on studies of related paramyxoviruses, the M protein is involved in assembling the RNP at the plasma membrane in regions containing the HN and F glycoproteins, presumably via interacting with the carboxy-terminal tail portion of the RNP-associated N protein and the cy...

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“…These structures are built of one or two domains that have similar β-sandwich folds, suggesting gene duplication during evolution. The dimers of paramyxovirus M protein (approximately 40 kDa) can form a grid-like array on the inner surface of the viral membrane (15)(16)(17)(18), and probably interact with both the cytoplasmic tails of the HN and F glycoproteins (19,20) as well as the nucleocapsid (21)(22)(23)(24) to initiate virus assembly and budding (20). The NDV M protein has greater than 20% sequence identity with other paramyxovirus M proteins including measles, mumps, and the parainfluenza viruses.…”

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confidence: 99%

Structure and assembly of a paramyxovirus matrix protein

Battisti

Meng

Winkler

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

126

141

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Many pleomorphic, lipid-enveloped viruses encode matrix proteins that direct their assembly and budding, but the mechanism of this process is unclear. We have combined X-ray crystallography and cryoelectron tomography to show that the matrix protein of Newcastle disease virus, a paramyxovirus and relative of measles virus, forms dimers that assemble into pseudotetrameric arrays that generate the membrane curvature necessary for virus budding. We show that the glycoproteins are anchored in the gaps between the matrix proteins and that the helical nucleocapsids are associated in register with the matrix arrays. About 90% of virions lack matrix arrays, suggesting that, in agreement with previous biological observations, the matrix protein needs to dissociate from the viral membrane during maturation, as is required for fusion and release of the nucleocapsid into the host's cytoplasm. Structure and sequence conservation imply that other paramyxovirus matrix proteins function similarly.

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The C-Terminal End of Parainfluenza Virus 5 NP Protein Is Important for Virus-Like Particle Production and M-NP Protein Interaction

Cited by 24 publications

References 54 publications

Matrix Proteins of Nipah and Hendra Viruses Interact with Beta Subunits of AP-3 Complexes

Matrix Proteins of Nipah and Hendra Viruses Interact with Beta Subunits of AP-3 Complexes

Gene-Specific Contributions to Mumps Virus Neurovirulence and Neuroattenuation

Structure and assembly of a paramyxovirus matrix protein

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