Tyrosine residues in the cytoplasmic domains affect sorting and fusion activity of the Nipah virus glycoproteins in polarized epithelial cells.
The highly pathogenic Nipah virus (NiV) is aerially transmitted and causes a systemic infection after entering the respiratory tract. Airway epithelia are thus important targets in primary infection. Furthermore, virus replication in the mucosal surfaces of the respiratory or urinary tract in later phases of infection is essential for virus shedding and transmission. So far, the mechanisms of NiV replication in epithelial cells are poorly elucidated. In the present study, we provide evidence that bipolar targeting of the two NiV surface glycoproteins G and F is of biological importance for fusion in polarized epithelia. We demonstrate that infection of polarized cells induces focus formation, with both glycoproteins located at lateral membranes of infected cells adjacent to uninfected cells. Supporting the idea of a direct spread of infection via lateral cell-to-cell fusion, we could identify basolateral targeting signals in the cytoplasmic domains of both NiV glycoproteins. Tyrosine 525 in the F protein is part of an endocytosis signal and is also responsible for basolateral sorting. Surprisingly, we identified a dityrosine motif at position 28/29 in the G protein, which mediates polarized targeting. A dileucine motif predicted to function as sorting signal is not involved. Mutation of the targeting signal in one of the NiV glycoproteins prevented the fusion of polarized cells, suggesting that basolateral or bipolar F and G expression facilitates the spread of NiV within epithelial cell monolayers, thereby contributing to efficient virus spread in mucosal surfaces in early and late phases of infection.
Weise
Carolin
C
Institute of Virology, Philipps University of Marburg, Marburg, Germany.
Erbar
Stephanie
S
Lamp
Boris
B
Vogt
Carola
C
Diederich
Sandra
S
Maisner
Andrea
A
D020816:Amino Acid Motifs; D000818:Animals; D002459:Cell Fusion; D004705:Endocytosis; D004847:Epithelial Cells / Q000821:virology*; D006801:Humans; D045405:Nipah Virus / Q000502:physiology*; D014443:Tyrosine / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000502:physiology*; D053586:Virus Internalization; D014779:Virus Replication
C510755:F protein, Nipah virus; D014759:Viral Envelope Proteins; C116204:attachment protein G; D014443:Tyrosine