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Influenza A Virus Hemagglutinin is Required for the Assembly of Viral Components Including Bundled vRNPs at the Lipid Raft

View Article: PubMed Central - PubMed

ABSTRACT

The influenza glycoproteins, hemagglutinin (HA) and neuraminidase (NA), which are associated with the lipid raft, have the potential to initiate virion budding. However, the role of these viral proteins in infectious virion assembly is still unclear. In addition, it is not known how the viral ribonucleoprotein complex (vRNP) is tethered to the budding site. Here, we show that HA is necessary for the efficient progeny virion production and vRNP packaging in the virion. We also found that the level of HA does not affect the bundling of the eight vRNP segments, despite reduced virion production. Detergent solubilization and a subsequent membrane flotation analysis indicated that the accumulation of nucleoprotein, viral polymerases, NA, and matrix protein 1 (M1) in the lipid raft fraction was delayed without HA. Based on our results, we inferred that HA plays a role in the accumulation of viral components, including bundled vRNPs, at the lipid raft.

No MeSH data available.


Related in: MedlinePlus

Decrease in virion production from cells infected with HAstop virus at both early and late phase of infection. MDCK cells were infected with WT and HAstop viruses. The supernatant of infected cells was collected at 15 hpi, and cells were suspended in MEM. The supernatant was also collected at 24 hpi. The virions were concentrated from the supernatant, and NP and M1 were detected by western blotting.
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viruses-08-00249-f005: Decrease in virion production from cells infected with HAstop virus at both early and late phase of infection. MDCK cells were infected with WT and HAstop viruses. The supernatant of infected cells was collected at 15 hpi, and cells were suspended in MEM. The supernatant was also collected at 24 hpi. The virions were concentrated from the supernatant, and NP and M1 were detected by western blotting.

Mentions: Our results show that the virion production is decreased and the association of viral components with lipid rafts is delayed in cells infected with HAstop virus. To analyze the relation between the decrease in virion production and the amount of viral proteins associated with lipid rafts in cells infected with HAstop, the number of virions in the supernatant from cells infected with WT and HAstop virus between 15 and 24 hpi was determined. The amounts of NP and M1 in the supernatant from cells infected with HAstop virus both between 1 and 15 hpi and between 15 and 24 hpi were decreased compared to that from cells infected with WT virus (Figure 5). This result suggests that the virion production is decreased from cells infected with HAstop virus even when the number of viral components associated with lipid rafts is comparable to that in cells infected with WT virus.


Influenza A Virus Hemagglutinin is Required for the Assembly of Viral Components Including Bundled vRNPs at the Lipid Raft
Decrease in virion production from cells infected with HAstop virus at both early and late phase of infection. MDCK cells were infected with WT and HAstop viruses. The supernatant of infected cells was collected at 15 hpi, and cells were suspended in MEM. The supernatant was also collected at 24 hpi. The virions were concentrated from the supernatant, and NP and M1 were detected by western blotting.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5035963&req=5

viruses-08-00249-f005: Decrease in virion production from cells infected with HAstop virus at both early and late phase of infection. MDCK cells were infected with WT and HAstop viruses. The supernatant of infected cells was collected at 15 hpi, and cells were suspended in MEM. The supernatant was also collected at 24 hpi. The virions were concentrated from the supernatant, and NP and M1 were detected by western blotting.
Mentions: Our results show that the virion production is decreased and the association of viral components with lipid rafts is delayed in cells infected with HAstop virus. To analyze the relation between the decrease in virion production and the amount of viral proteins associated with lipid rafts in cells infected with HAstop, the number of virions in the supernatant from cells infected with WT and HAstop virus between 15 and 24 hpi was determined. The amounts of NP and M1 in the supernatant from cells infected with HAstop virus both between 1 and 15 hpi and between 15 and 24 hpi were decreased compared to that from cells infected with WT virus (Figure 5). This result suggests that the virion production is decreased from cells infected with HAstop virus even when the number of viral components associated with lipid rafts is comparable to that in cells infected with WT virus.

View Article: PubMed Central - PubMed

ABSTRACT

The influenza glycoproteins, hemagglutinin (HA) and neuraminidase (NA), which are associated with the lipid raft, have the potential to initiate virion budding. However, the role of these viral proteins in infectious virion assembly is still unclear. In addition, it is not known how the viral ribonucleoprotein complex (vRNP) is tethered to the budding site. Here, we show that HA is necessary for the efficient progeny virion production and vRNP packaging in the virion. We also found that the level of HA does not affect the bundling of the eight vRNP segments, despite reduced virion production. Detergent solubilization and a subsequent membrane flotation analysis indicated that the accumulation of nucleoprotein, viral polymerases, NA, and matrix protein 1 (M1) in the lipid raft fraction was delayed without HA. Based on our results, we inferred that HA plays a role in the accumulation of viral components, including bundled vRNPs, at the lipid raft.

No MeSH data available.


Related in: MedlinePlus