Novel Phosphoprotein-interacting Region in Nipah Virus Nucleocapsid Protein and Its Involvement in Viral Replication

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2010 Jul 30

PMID 20668089

Citations 20

Authors

Mio Omi-Furutani

Misako Yoneda

Kentaro Fujita

Fusako Ikeda

Chieko Kai

Affiliations

Soon will be listed here.

Abstract

The interaction of Nipah virus (NiV) nucleocapsid (N) protein with phosphoprotein (P) during nucleocapsid assembly is the essential process in the viral life cycle, since only the encapsidated RNA genome can be used for replication. To identify the region responsible for N-P interaction, we utilized fluorescent protein tags to visualize NiV N and P proteins in live cells and analyzed their cellular localization. N protein fused to monomeric enhanced cyan fluorescence protein (N-ECFP) exhibited a dotted pattern in transfected cells, while P protein fused to monomeric red fluorescent protein (P-mRFP) showed diffuse distribution. When the two proteins were coexpressed, P-mRFP colocalized with N-ECFP dots. N-ECFP mutants with serial amino acid deletions were generated to search for the region(s) responsible for this N-P colocalization. We found that, in addition to the 467- to 496-amino-acid (aa) region reported previously, aa 135 to 146 were responsible for the N-P colocalization. The residues crucial for N-P interaction were further investigated by introducing alanine substitutions into the untagged N protein. Alanine scanning in the region of aa 135 to 146 has revealed that there are distinct regions essential for the interaction of N-P and the function of N. This is the first study to visualize Nipah viral proteins in live cells and to assess the essential domain of N protein for the interaction with P protein.

Citing Articles

In silico designing of an epitope-based peptide vaccine cocktail against Nipah virus: an Indian population-based epidemiological study.

Chaudhuri D, Majumder S, Datta J, Giri K Arch Microbiol. 2023; 205(12):380.

PMID: 37955744 DOI: 10.1007/s00203-023-03717-3.

Genomic characterization, transcriptome analysis, and pathogenicity of the Nipah virus (Indian isolate).

Mohandas S, Shete A, Sarkale P, Kumar A, Mote C, Yadav P Virulence. 2023; 14(1):2224642.

PMID: 37312405 PMC: 10281463. DOI: 10.1080/21505594.2023.2224642.

Nipah Virus Impairs Autocrine IFN Signaling by Sequestering STAT1 and STAT2 into Inclusion Bodies.

Becker N, Maisner A Viruses. 2023; 15(2).

PMID: 36851768 PMC: 9967463. DOI: 10.3390/v15020554.

The Nucleocapsid of Paramyxoviruses: Structure and Function of an Encapsidated Template.

Bloyet L Viruses. 2021; 13(12).

PMID: 34960734 PMC: 8708338. DOI: 10.3390/v13122465.

Zinc and Copper Ions Differentially Regulate Prion-Like Phase Separation Dynamics of Pan-Virus Nucleocapsid Biomolecular Condensates.

Monette A, Mouland A Viruses. 2020; 12(10).

PMID: 33081049 PMC: 7589941. DOI: 10.3390/v12101179.

References

RYAN K, Portner A, Murti K . Antibodies to paramyxovirus nucleoproteins define regions important for immunogenicity and nucleocapsid assembly. Virology. 1993; 193(1):376-84. DOI: 10.1006/viro.1993.1134. View

Bankamp B, Horikami S, Thompson P, Huber M, Billeter M, Moyer S . Domains of the measles virus N protein required for binding to P protein and self-assembly. Virology. 1996; 216(1):272-7. DOI: 10.1006/viro.1996.0060. View

Curran J, Marq J, Kolakofsky D . An N-terminal domain of the Sendai paramyxovirus P protein acts as a chaperone for the NP protein during the nascent chain assembly step of genome replication. J Virol. 1995; 69(2):849-55. PMC: 188651. DOI: 10.1128/JVI.69.2.849-855.1995. View

Slack M, Easton A . Characterization of the interaction of the human respiratory syncytial virus phosphoprotein and nucleocapsid protein using the two-hybrid system. Virus Res. 1998; 55(2):167-76. DOI: 10.1016/s0168-1702(98)00042-2. View

Randall R, Bermingham A . NP:P and NP:V interactions of the paramyxovirus simian virus 5 examined using a novel protein:protein capture assay. Virology. 1996; 224(1):121-9. DOI: 10.1006/viro.1996.0513. View

Fooks A, Stephenson J, Warnes A, Dowsett A, Rima B, Wilkinson G . Measles virus nucleocapsid protein expressed in insect cells assembles into nucleocapsid-like structures. J Gen Virol. 1993; 74 ( Pt 7):1439-44. DOI: 10.1099/0022-1317-74-7-1439. View

Chua K, Bellini W, Rota P, Harcourt B, Tamin A, Lam S . Nipah virus: a recently emergent deadly paramyxovirus. Science. 2000; 288(5470):1432-5. DOI: 10.1126/science.288.5470.1432. View

Chan Y, Koh C, Lam S, Wang L . Mapping of domains responsible for nucleocapsid protein-phosphoprotein interaction of Henipaviruses. J Gen Virol. 2004; 85(Pt 6):1675-1684. DOI: 10.1099/vir.0.19752-0. View

Zhao H, Banerjee A . Interaction between the nucleocapsid protein and the phosphoprotein of human parainfluenza virus 3. Mapping of the interacting domains using a two-hybrid system. J Biol Chem. 1995; 270(21):12485-90. DOI: 10.1074/jbc.270.21.12485. View

10.

Sugimoto K, Uema M, Sagara H, Tanaka M, Sata T, Hashimoto Y . Simultaneous tracking of capsid, tegument, and envelope protein localization in living cells infected with triply fluorescent herpes simplex virus 1. J Virol. 2008; 82(11):5198-211. PMC: 2395178. DOI: 10.1128/JVI.02681-07. View

11.

Curran J, Homann H, Buchholz C, Rochat S, Neubert W, Kolakofsky D . The hypervariable C-terminal tail of the Sendai paramyxovirus nucleocapsid protein is required for template function but not for RNA encapsidation. J Virol. 1993; 67(7):4358-64. PMC: 237806. DOI: 10.1128/JVI.67.7.4358-4364.1993. View

12.

Ong S, Yusoff K, Kho C, Abdullah J, Tan W . Mutagenesis of the nucleocapsid protein of Nipah virus involved in capsid assembly. J Gen Virol. 2009; 90(Pt 2):392-397. DOI: 10.1099/vir.0.005710-0. View

13.

Yoneda M, Guillaume V, Ikeda F, Sakuma Y, Sato H, Wild T . Establishment of a Nipah virus rescue system. Proc Natl Acad Sci U S A. 2006; 103(44):16508-13. PMC: 1618306. DOI: 10.1073/pnas.0606972103. View

14.

Halpin K, Young P, Field H, MacKenzie J . Newly discovered viruses of flying foxes. Vet Microbiol. 1999; 68(1-2):83-7. DOI: 10.1016/s0378-1135(99)00063-2. View

15.

Chenik M, Chebli K, Gaudin Y, Blondel D . In vivo interaction of rabies virus phosphoprotein (P) and nucleoprotein (N): existence of two N-binding sites on P protein. J Gen Virol. 1994; 75 ( Pt 11):2889-96. DOI: 10.1099/0022-1317-75-11-2889. View

16.

Howard M, Wertz G . Vesicular stomatitis virus RNA replication: a role for the NS protein. J Gen Virol. 1989; 70 ( Pt 10):2683-94. DOI: 10.1099/0022-1317-70-10-2683. View

17.

Liston P, Batal R, DiFlumeri C, Briedis D . Protein interaction domains of the measles virus nucleocapsid protein (NP). Arch Virol. 1997; 142(2):305-21. DOI: 10.1007/s007050050078. View

18.

Hsu V, Hossain M, Parashar U, Ali M, Ksiazek T, Kuzmin I . Nipah virus encephalitis reemergence, Bangladesh. Emerg Infect Dis. 2005; 10(12):2082-7. PMC: 3323384. DOI: 10.3201/eid1012.040701. View

19.

Lu B, Brazas R, Ma C, Kristoff T, Cheng X, Jin H . Identification of temperature-sensitive mutations in the phosphoprotein of respiratory syncytial virus that are likely involved in its interaction with the nucleoprotein. J Virol. 2002; 76(6):2871-80. PMC: 135989. DOI: 10.1128/jvi.76.6.2871-2880.2002. View

20.

Halpin K, Bankamp B, Harcourt B, Bellini W, Rota P . Nipah virus conforms to the rule of six in a minigenome replication assay. J Gen Virol. 2004; 85(Pt 3):701-707. DOI: 10.1099/vir.0.19685-0. View