A Leucine Residue in the C Terminus of Human Parainfluenza Virus Type 3 Matrix Protein Is Essential for Efficient Virus-Like Particle and Virion Release

Zhang, Guangyuan; Zhang, Shengwei; Ding, Beichen; Yang, Xiaodan; Chen, Longyun; Yan, Qin; Jiang, Yanliang; Zhong, Yi; Chen, Mingzhou

doi:10.1128/jvi.01485-14

Cited by 20 publications

(33 citation statements)

References 66 publications

(62 reference statements)

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“…While the fusogenicity of NiVeG⌬M was slightly enhanced, replication kinetics was significantly impaired. Together with the release of large amounts of viral and cellular proteins and the severely reduced infectivity and stability, our data indicate that the M protein is essential for proper NiV particle assembly, consistent with its role in other paramyxoviruses (21,(31)(32)(33).…”

Section: Discussionsupporting

confidence: 54%

“…While some attempts to generate members of the Paramyxoviridae family without complementation of functional M proteins were not successful (31)(32)(33), M protein-deleted measles (MV⌬M) and respiratory syncytial (M-null HRSV) viruses have been recovered (20,21). While M-null HRSV was completely defective in virus budding (21), limited amounts of infectious MV⌬M were found in the supernatant (20), indicating a varying importance of the M protein contribution among different genera.…”

Section: Discussionmentioning

confidence: 99%

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Nipah Virus Matrix Protein Influences Fusogenicity and Is Essential for Particle Infectivity and Stability

Dietzel

Kolesnikova

Sawatsky

et al. 2016

J Virol

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Nipah virus (NiV) causes fatal encephalitic infections in humans. To characterize the role of the matrix (M) protein in the viral life cycle, we generated a reverse genetics system based on NiV strain Malaysia. Using an enhanced green fluorescent protein (eGFP)-expressing M protein-deleted NiV, we observed a slightly increased cell-cell fusion, slow replication kinetics, and significantly reduced peak titers compared to the parental virus. While increased amounts of viral proteins were found in the supernatant of cells infected with M-deleted NiV, the infectivity-to-particle ratio was more than 100-fold reduced, and the particles were less thermostable and of more irregular morphology. Taken together, our data demonstrate that the M protein is not absolutely required for the production of cell-free NiV but is necessary for proper assembly and release of stable infectious NiV particles. IMPORTANCEHenipaviruses cause a severe disease with high mortality in human patients. Therefore, these viruses can be studied only in biosafety level 4 (BSL-4) laboratories, making it more challenging to characterize their life cycle. Here we investigated the role of the Nipah virus matrix protein in virus-mediated cell-cell fusion and in the formation and release of newly produced particles. We found that even though low levels of infectious viruses are produced in the absence of the matrix protein, it is required for the release of highly infectious and stable particles. Fusogenicity of matrixless viruses was slightly enhanced, further demonstrating the critical role of this protein in different steps of Nipah virus spread.

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Section: Discussionsupporting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Nipah Virus Matrix Protein Influences Fusogenicity and Is Essential for Particle Infectivity and Stability

Dietzel

Kolesnikova

Sawatsky

et al. 2016

J Virol

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“…The plasmids carrying HA-M, HA-M L305A , HA-P, Myc-M, Myc-M L305A , Myc-N, and N-Flag have been described previously (15). pOCUS-HPIV3 was used as a template for PCR amplification in the genetic manipulations.…”

Section: Cells and Virus Cells (293t And Llc-mk2mentioning

confidence: 99%

“…Monolayers of MK2 cells in 6-well plates were grown to 50 to 60% confluence and infected with wildtype HPIV3 or HPIV3 expressing M L305A (HPIV3-M L305A ) at an MOI of 0.001. Then, the infection medium was removed and replaced with fresh medium containing 4% FBS, and the supernatant was sampled every 12 h for 4 days prior to determination of the infectious virus titer, which was performed by standard plaque assay on MK2 cells as described previously (15). After the viral titers were calculated, growth curves were determined by using the software GraphPad Prism (version 5.0).…”

Section: Cells and Virus Cells (293t And Llc-mk2mentioning

confidence: 99%

“…In contrast, the M proteins of other NSVs, such as mumps virus (6) and parainfluenza virus type 5 (PIV5) (14), require an accessory protein, e.g., F or N, for maximum VLP release efficiency. Previously, we also demonstrated that the M protein of human parainfluenza virus type 3 (HPIV3) alone in mammalian cells could lead to the formation and release of enveloped VLPs (15), which are morphologically similar to virions.…”

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confidence: 99%

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Interaction of Human Parainfluenza Virus Type 3 Nucleoprotein with Matrix Protein Mediates Internal Viral Protein Assembly

Zhang

Zhong

Qin

et al. 2016

J Virol

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Human parainfluenza virus type 3 (HPIV3) belongs to the Paramyxoviridae family. Its three internal viral proteins, the nucleoprotein (N), the phosphoprotein (P), and the polymerase (L), form the ribonucleoprotein (RNP) complex, which encapsidates the viral genome and associates with the matrix protein (M) for virion assembly. We previously showed that the M protein expressed alone is sufficient to assemble and release virus-like particles (VLPs) and a mutant with the L305A point mutation in the M protein (M L305A ) has a VLP formation ability similar to that of wild-type M protein. In addition, recombinant HPIV3 (rHPIV3) containing the M L305A mutation (rHPIV3-M L305A ) could be successfully recovered. In the present study, we found that the titer of rHPIV3-M L305A was at least 10-fold lower than the titer of rHPIV3. Using VLP incorporation and coimmunoprecipitation assays, we found that VLPs expressing the M protein (M-VLPs) can efficiently incorporate N and P via an N-M or P-M interaction and M L305A -VLPs had an ability to incorporate P via a P-M interaction similar to that of M-VLPs but were unable to incorporate N and no longer interacted with N. Furthermore, we found that the incorporation of P into M L305A -VLPs but not M-VLPs was inhibited in the presence of N. In addition, we provide evidence that the C-terminal region of P is involved in its interaction with both N and M and N binding to the C-terminal region of P inhibits the incorporation of P into M L305A -VLPs. Our findings provide new molecular details to support the idea that the N-M interaction and not the P-M interaction is critical for packaging N and P into infectious viral particles. IMPORTANCEHuman parainfluenza virus type 3 (HPIV3) is a nonsegmented, negative-sense, single-stranded RNA virus that belongs to the Paramyxoviridae family and can cause lower respiratory tract infections in infants and young children as well as elderly or immunocompromised individuals. However, no effective vaccine has been developed or licensed. We used virus-like particle (VLP) incorporation and coimmunoprecipitation assays to determine how the M protein assembles internal viral proteins. We demonstrate that both nucleoprotein (N) and phosphoprotein (P) can incorporate into M-VLPs and N inhibits the M-P interaction via the binding of N to the C terminus of P. We also provide additional evidence that the N-M interaction but not the P-M interaction is critical for the regulation of HPIV3 assembly. Our studies provide a more complete characterization of HPIV3 virion assembly and substantiation that N interaction with M regulates internal viral organization. Human parainfluenza virus type 3 (HPIV3) is a negativestrand RNA virus (NSV) that belongs to the Paramyxoviridae family and often causes lower respiratory tract infections in infants and young children. The HPIV3 genome consists of 6 open reading frames that encode 6 structural proteins: the nucleoprotein (N), phosphoprotein (P), RNA-dependent RNA polymerase (L), matrix protein (M), and two glycoprotei...

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The interplay between pathogens and Atg8 family proteins: thousand‐faced interactions

2021

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Autophagy is an intracellular degradation and recycling process that can also remove pathogenic intracellular bacteria and viruses from within cells (referred to as xenophagy) and activate the adaptive immune responses. But autophagy—especially Atg proteins including Atg8 family members—can also have proviral and probacterial effects. In this review, we summarize known interactions of bacterial, parasitic, and viral proteins with Atg8 family proteins and the outcome of these interactions on pathogen replication, autophagy, or mitophagy. We discuss the value of prediction software and the research methodology in the study of pathogen protein‐Atg8 family protein interactions, with selected examples of potential LC3‐interacting region motif‐containing SARS‐CoV‐2 proteins.

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A Leucine Residue in the C Terminus of Human Parainfluenza Virus Type 3 Matrix Protein Is Essential for Efficient Virus-Like Particle and Virion Release

Cited by 20 publications

References 66 publications

Nipah Virus Matrix Protein Influences Fusogenicity and Is Essential for Particle Infectivity and Stability

Nipah Virus Matrix Protein Influences Fusogenicity and Is Essential for Particle Infectivity and Stability

Interaction of Human Parainfluenza Virus Type 3 Nucleoprotein with Matrix Protein Mediates Internal Viral Protein Assembly

The interplay between pathogens and Atg8 family proteins: thousand‐faced interactions

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