Residue 41 of the Eurasian avian-like swine influenza a virus matrix protein modulates virion filament length and efficiency of contact transmission.
Position 41 of the influenza A virus matrix protein encodes a highly conserved alanine in human and avian lineages. Nonetheless, strains of the Eurasian avian-like swine (Easw) lineage contain a change at this position: position 41 of A/swine/Spain/53207/04 (H1N1) (SPN04) encodes a proline. To assess the impact of this naturally occurring polymorphism on viral fitness, we utilized reverse genetics to produce recombinant viruses encoding wild-type M1 41P (rSPN04-P) and consensus 41A (rSPN04-A) residues. Relative to rSPN04-A, rSPN04-P virus displayed reduced growth in vitro. In the guinea pig model, rSPN04-P was transmitted to fewer contact animals than rSPN04-A and failed to infect guinea pigs that received a low-dose inoculum. Moreover, the P41A change altered virion morphology, reducing the number and length of filamentous virions, as well as reducing the neuraminidase activity of virions. The lab-adapted human isolate, A/PR/8/34 (H1N1) (PR8), is nontransmissible in the guinea pig model, making it a useful background in which to identify certain viral factors that enhance transmissibility. We assessed transmission in the context of single-, double-, and triple-reassortant viruses between PR8 and SPN04; PR8/SPN04 M, PR8/SPN04 M+NA, and PR8/SPN04 M+NA+HA, encoding either matrix 41 A or P, were generated. In each case, the virus possessing 41P transmitted less well than the corresponding 41A-encoding virus. In summary, we have identified a naturally occurring mutation in the influenza A virus matrix protein that impacts transmission efficiency and can alter virion morphology and neuraminidase activity.


OBJECTIVE
We have developed a practical model for examining the genetics underlying transmissibility of the Eurasian avian-like swine lineage viruses, which contributed M and NA segments to the 2009 pandemic strain. Here, we use our system to investigate the impact on viral fitness of a naturally occurring polymorphism at matrix (M1) position 41 in an Easw isolate. Position 41 has been implicated previously in adaptation to laboratory substrates and to mice. Here we show that the polymorphism at M1 41 has a limited effect on growth in vitro but changes the morphology of the virus and impacts growth and transmission in the guinea pig model.
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA Microbiology and Molecular Genetics Graduate Program, Emory University, Atlanta, Georgia, USA.
Patricia J
PJ
Campbell
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA.
Constantinos S
CS
Kyriakis
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA Microbiology and Molecular Genetics Graduate Program, Emory University, Atlanta, Georgia, USA.
Nicolle
N
Marshall
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA.
Suganthi
S
Suppiah
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA Microbiology and Molecular Genetics Graduate Program, Emory University, Atlanta, Georgia, USA.
Jill
J
Seladi-Schulman
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA.
Shamika
S
Danzy
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA.
Anice C
AC
Lowen
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA john.steel@emory.edu.
John
J
Steel
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D015151:Immunoblotting; D053118:Influenza A Virus, H1N1 Subtype / Q000502:physiology*; D051379:Mice; D009154:Mutation / Q000235:genetics; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism*; D009976:Orthomyxoviridae Infections / Q000635:transmission* / Q000821:virology; D016865:Reassortant Viruses / Q000502:physiology*; D013552:Swine; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000502:physiology*
C031540:M-protein, influenza virus; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C487630:NA protein, influenza A virus; D009439:Neuraminidase