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Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein
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Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Published on: April 21, 2015

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Conserved methionine 165 of matrix protein contributes to the nuclear import and is essential for influenza A virus

Petra Švančarová1, Tatiana Betáková2

  • 1Biomedical Research Center - Slovaks Academy of Sciences, Institute of Virology, Bratislava, Slovak Republic.

Virology Journal
|December 5, 2018
PubMed
Abstract

Insights

Methionine 165 in influenza matrix protein (M1) is crucial for virus replication. The conserved RQMV motif, particularly M165, is vital for nuclear import of viral proteins, impacting influenza virus assembly and infection.

Area of Science:

  • Virology
  • Molecular Biology

Background:

  • The influenza matrix protein (M1) is essential for virus assembly and infection, located beneath the viral membrane.
  • M1 protein comprises N-domain and C-domain, linked by a loop region containing the conserved RQMV motif.

Purpose of the Study:

  • To investigate the function of the conserved RQMV motif in the influenza virus life cycle.
  • To elucidate the role of specific amino acids within the RQMV motif in viral protein localization and replication.

Main Methods:

  • Site-directed mutagenesis was employed to create M1 mutants within the RQMV motif.
  • Rescuing mutant viruses using reverse genetics allowed for in-depth analysis.
  • Confocal microscopy was utilized to study the co-localization of M1 with key cellular proteins (NP, CLUH, CRM1) and the plasma membrane.

Main Results:

  • Mutations at R163, Q164, and V166 did not affect viral phenotype, producing wild-type virus.
  • A single M165A substitution proved lethal for viral rescue, significantly impacting M1 and nucleoprotein (NP) distribution.
  • M165A and NP showed reduced nuclear distribution (p<0.05 and p<0.001, respectively).
  • M165A co-localized with CLUH in the cytoplasm and perinuclear region, with restricted transport through the nuclear membrane.

Conclusions:

  • Methionine 165 (M165) is indispensable for influenza virus replication.
  • The RQMV motif plays a critical role in the nuclear import of viral proteins, essential for the influenza virus life cycle.
Keywords:
CLUHInfluenza a virusM1 proteinNP proteinReverse genetic

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