Highly dynamic microtubules improve the effectiveness of early stages of human influenza A/NWS/33 virus infection in LLC-MK2 cells.
BACKGROUND
This study aims to investigate the role of microtubule dynamics in the initiation of NWS/33 human influenza A (NWS) virus infection in MDCK and LLC-MK2 mammalian kidney cells. We previously demonstrated a host-dependent role of the actin cytoskeleton in inducing restriction during the early phases of NWS infection. Furthermore, we showed the differential infectious entry of NWS virus in the above mentioned cell models.


RESULTS
By first employing a panel of microtubule-modulators, we evidenced that microtubule-stabilization negatively interferes with NWS replication in LLC-MK2 but not in MDCK cells. Conversely, microtubule-depolymerization improves NWS growth in LLC-MK2 but not in the MDCK model. By using immunofluorescence labelling and Western blotting analyses upon NWS infection in mammalian kidney cells, it was observed that the occurrence of alpha-tubulin hyperacetylation--a post-translational modified form suggestive of stable microtubules--was significantly delayed in LLC-MK2 when compared to MDCK cells. Furthermore, mock-infected LLC-MK2 cells were shown to have higher levels of both acetylated alpha-tubulin and microtubule-associated protein 4 (MAP4), the latter being essential for the maintenance of normal microtubule polymer levels in interphase epithelial cells. Finally, to obtain highly dynamic microtubules in LLC-MK2 cells, we knocked down the expression of MAP4 by using a RNA-mediated RNA interference approach. The results evidenced that MAP4 silencing improves NWS growth in LLC-MK2 cells.


CONCLUSIONS
By evidencing the cell type-dependent regulatory role of microtubule dynamics on NWS replication in mammalian kidney cells, we demonstrated that microtubule-stabilization represents a restriction factor for the initiation of NWS infection in LLC-MK2 but not in MDCK cells.
Section of Microbiology, Department of Pathology and Laboratory Medicine, University of Parma, Parma, Italy. flora.deconto@unipr.it
Flora
F
De Conto
Enrica
E
Di Lonardo
Maria Cristina
MC
Arcangeletti
Carlo
C
Chezzi
Maria Cristina
MC
Medici
Adriana
A
Calderaro
D000107:Acetylation; D000818:Animals; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D004285:Dogs; D005786:Gene Expression Regulation; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus / Q000502:physiology*; D008253:Macaca mulatta; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology; D009928:Organ Specificity / Q000235:genetics; D017239:Paclitaxel / Q000494:pharmacology; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D034622:RNA Interference; D013552:Swine; D014404:Tubulin / Q000378:metabolism; D050257:Tubulin Modulators / Q000494:pharmacology; D014779:Virus Replication / Q000187:drug effects
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D050257:Tubulin Modulators; C483468:hemagglutinin, human influenza A virus; D017239:Paclitaxel; D015739:Nocodazole